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Trade deal progress; Politburo reports to Xi; Demographics; Confucius Institutes
Bill Bishop Feb 28, 2019 4 Comment Share
The official spin cycle is revving up to sell any trade agreement with China as a huge win, even though I am still hearing the mooted deal is far from what the US originally wanted. One of the PR talking points from the Trump Administration seems to be that this is a "process" and the US can't expect to get everything in just one deal.
The Chinese side is messaging that their concessions are huge and they may be difficult for Xi to accept, or to implement in the near term. So while clearly there is momentum towards a deal the details are not done, and probably will not be until Trump and Xi meet in late March.
Kim Jong-un will return home to North Korea through China this weekend. There is speculation that he may stop in Beijing this time to meet with Xi but so far nothing is confirmed. The Chinese side wanted more progress from this second summit and would very much like to officially loosen sanctions on the DPRK.
Perhaps Beijing will see Trump's decision to walk away from the Kim summit without a deal as a sign that he is willing to walk from the trade deal under discussion, but given the upcoming election cycle US domestic politics are increasingly unfavorable for President Trump to continue the trade war and so I am not sure how much of a signaling effect it will really have.
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1. US-China trade
Kudlow's ebullience was remarkable - Larry Kudlow says US-China trade deal talks made 'fantastic' progress last week - CNBC:
"Last week was fantastic," Kudlow told CNBC's "Squawk on the Street." "We're making great headway on nontariff barriers and tariffs regarding various commodities such as soybeans and energy and beef. We have mechanisms with regard to enforcement, which is — I think — unparalleled."
"The progress has been terrific," Kudlow added. But "we have to hear from the Chinese side. We have to hear from President Xi Jinping, of course. I think we're headed for a remarkable, historic deal."
China deal remains close - POLITICO:
MM is told by a senior administration official that there isn't much tension between Lighthizer and Trump, who speak nearly every day. This person described the China deal as largely complete, currently running to around 140 pages and including significant enforcement mechanisms. If the Chinese are found to be in violation of the deal, the U.S. would be able to re-impose tariffs. The Chinese would not be allowed to retaliate.
Waiting on Xi — The enforcement structures would be a big leap for the Chinese. And it's still unclear whether Chinese President Xi Jinping will ultimately agree to them. So a deal could still fail. But the senior administration official told MM that if Xi accedes to what his negotiators have put on paper then there will likely be a signing event at Mar-a-Lago at the end of March.
Lighthizer's reality check on China talks - POLITICO:
Lighthizer really wants this to be bipartisan. Throughout the hearing, he repeatedly praised House Speaker Nancy Pelosi for her opposition to the U.S. trade bill that accompanied China's entry into the World Trade Organization, arguing the United States would be in a better position if her view had prevailed.
"These things have to be bipartisan. They cannot be partisan," Lighthizer said. "This is just different than a lot of other stuff we deal with."
— A currency deal isn't a done deal yet, despite Trump's comments last week signaling otherwise. "There's no agreement on anything until there's an agreement on everything. But the reality is we have spent a lot of time on currency and it'll be enforceable," Lighthizer said
USTR to suspend China tariff hike 'until further notice' | Reuters:
In accordance with the president's direction, the Office of the United States Trade Representative will publish a notice in the Federal Register this week to suspend the scheduled tariff increase until further notice
Trump trade official calls for strict policing of U.S.-China deal - The Washington Post:
Under the U.S. proposal, working-level officials would meet each month to consider complaints about Chinese practices from American businesses, including ones submitted anonymously to avoid stirring Beijing's ire, Lighthizer said.
Those that can't be resolved will pass to higher level officials meeting quarterly and, if necessary, to Lighthizer and his Chinese counterpart, Vice Premier Liu He, who would confab twice a year.
Mnuchin says US-China trade deal will have real structural commitments, but still more work to do:
"The deal is not done yet but we have made a lot of progress" Mnuchin told CNBC's Hadley Gamble in an interview that aired on "Squawk on the Street" Thursday. "We still have more work to do, and we hope to make progress this month."
Opening Statement of USTR Robert Lighthizer to the House Ways and Means Committee | United States Trade Representative:
I would just like to read something briefly. Faced with the PNTR [Permanent Normal Trade Relations] vote in April of 2000, the Speaker said:
It is incumbent upon all of us in the public and private sectors to work for free and open trade with China that is real. The U.S.-China bilateral WTO agreement, however, is seriously deficient in substance, implementation, and enforcement. This issue is too important for our economy to be based on a pattern of broken promises, not proven performance. China can become a member of the WTO without Congress having to surrender its right to U.S.-China trade review annually. There is no reason why we should permanently surrender that leverage at this time.
I ask: if her position had prevailed, how different would things be right now? There are many other examples of bipartisan leadership, including a lot of people on this committee and I'm going to get into it and answer the questions if it's relevant.
U.S. Trade Representative Says China Deal Will Include Enforcement Mechanism - WSJ $$:
In his testimony, Mr. Lighthizer said any deal with China must have strong enforcement provisions to ensure Beijing honors the deal. Complaints of violations would be discussed in a series of consultations, he said—monthly by staffers, quarterly by vice ministers and twice-yearly by ministers of the two nations. That last would likely mean Mr. Lighthizer and Chinese Vice Premier Liu He, China's special envoy on trade.
US trade demands could take China years to achieve, says Beijing's ambassador to Washington | South China Morning Post:
Cui Tiankai, the Chinese ambassador to the US, also hit out at the US government's criminal action against Chinese telecoms company Huawei, calling it "very stupid" and "absurd".
Discussing recent trade negotiations, Cui told the South China Morning Post that while certain commitments such as purchases of US goods could be realised in a short time frame, other reforms that would have to go through China's legislative process could take anywhere from "five to 10 years" to accomplish.
One sign of the mood in DC - CPAC's new boogeyman: China - POLITICO:
Last year, the title of a panel at the annual Conservative Political Action Conference posed a simple question — "What is the Biggest Threat to the U.S.?" — and offered three options: China, Russia or rogue states like North Korea.
Now, the results are in. "China, the global menace," warns the title of one panel at this year's conference. "21st Century terminator: How China is using 5G and AI to take over the world," warns another. A third China-focused panel borrows its title from a book by Winston Churchill about the runup to World War II. The words "Russia," "Iran," "North Korea" and "terrorism" do not appear on this year's agenda.
2. Politburo members submit work reports to their boss
A reminder they all work for Xi, in case there were still doubts…
Senior CPC officials report work to CPC Central Committee, Xi - Xinhua:
Members of the Political Bureau of the Communist Party of China (CPC) Central Committee recently submitted reports on their work to the CPC Central Committee and General Secretary Xi Jinping in accordance with regulations, an official statement said Thursday.
After reading through their work reports, Xi urged them to fulfill their historical responsibilities given by the Party and the people, always preserve the political character and revolutionary spirit of Communists and devote themselves to public duty, said the statement.
Xi called on members to be enterprising, tackle tough issues head-on, maintain a strong sense of responsibility, uphold integrity and self-discipline, work diligently and set an example for the whole Party.
The members of the Political Bureau of the CPC Central Committee wrote their reports on their work over the last year, which included summaries of their work experiences, problems they found and improvements they did, as well as the plans and advice for their future work.
中央政治局同志向党中央和习近平总书记述职
China's ruling elite tell Xi they have 'strengthened education' of families | Reuters:
"Comrades of the Politburo strictly enforce the rules of honesty and self-discipline of party members and leading cadres ... and strengthen the education management of relatives and staff around them," the report added.
3. Huawei
Merkel seeking no-spying deal with China over Huawei issue: magazine | Reuters:
Chancellor Angela Merkel wants to agree a deal with China not to spy on each other so that Germany does not have to exclude Huawei on principle from building a 5G network in Germany, a magazine on Tuesday cited security sources as saying.
"Merkel's economic advisor Lars-Hendrik Roller was recently in China to negotiate about the agreement," WirtschaftsWoche cited security sources as saying.
German cyber security chief backs 5G 'no spy' deal over Huawei | Financial Times $$:
The remarks by Arne Schönbohm, head of the BSI, the German federal cyber security agency, suggest that Germany could formally allow Huawei to participate in the 5G roll-out if Chinese authorities give extra assurances on data security.
That would mark a setback for attempts by the US to persuade its allies to ban Huawei on the grounds that the Chinese state could use the company to conduct espionage or cyber sabotage.
The PRC Embassy in DC sent me a copy of this letter - 'Come and see us,' Huawei says in open letter to US media - China Daily:
"The US is a shining example of how to inspire passion for technological innovation and development. We too have been inspired by your history of creativity and hard work," said the letter, bearing the name Catherine Chen, a Director of the Board at Huawei, who is in charge of public and government affairs.
"I am writing to you in the hopes that we can come to understand each other better. In recent years, the US government has developed some misunderstandings about us. We would like to draw your attention to the facts," the letter said.
How much do Chinese people love Huawei? Just ask these cute singing children. - The Washington Post:
A music video featuring a group of kids performing a song called "Huawei the Beautiful" went viral on Chinese social media Wednesday.
The video was made by Zhoudan Kids' Singing Classroom, a production company based in Zhuhai in southern China, which posted the video on its WeChat social media account. It was then widely shared on Weibo, China's answer to Twitter.
The production company explained that it made the video as "an act of public benefit" and said that all children involved, and their parents, were real-life fans of Huawei.
RBC Said to Ask Bankers Avoid China Travel Amid Huawei Saga - Bloomberg:
The Toronto-based lender has asked its workers, especially those based in Canada, to skip business trips to China, one of the people said. The bank has requested that at least two Canada-based staff cancel or delay their travel to the country, according to the person, who asked not to be identified because the information is private.
The Canadian minister who could decide whether Huawei's Meng Wanzhou goes free | South China Morning Post:
According to section 14 of Canada's Extradition Act, Lametti has 30 days from when the US formally filed its extradition request to make a decision...
The Canadian Department of Justice confirmed on January 29 that officials had received a formal extradition request from the US. Assuming the request was filed on that date...
The decision is widely expected to be announced at the March 6 hearing.
Five Eyes countries should follow national interests rather than US opinions - Global Times:
The US has been trying to rope in the other four countries from the Five Eyes (FVEY) intelligence alliance, the UK, Canada, Australia, and New Zealand, to encircle China's Huawei Technologies. However, the UK National Cyber Security Center has recently determined that "there are ways to limit the risks from using Huawei in future 5G."
New Zealand also confirmed that Huawei hasn't been ruled out of from playing a role in the development of the nation's 5G network. FVEY once followed Washington's opinion, but now the UK and New Zealand are taking different stands.
4. PBoC official reiterates no credit surge coming
Central Bank Official Downplays Chances of Credit Binge - Caixin:
"(We) will fully take into account new changes in the economic and financial situation to fine-tune (policies), but we will do this in a measured manner and firmly avoid flooding the economy (with credit)," Wang Jingwu, head of the Financial Stability Bureau of the People's Bank of China (PBOC), said in an interview with the Financial News, a newspaper run by the monetary authority. The bureau is responsible for guarding against systemic risks in the financial system
"(We) will keep a close eye on and carry out a dynamic assessment of market liquidity and factors affecting it, and be vigilant against the formation of a 'liquidity delusion' and expectations of one-way (moves in the monetary policy) among market participants," he said.
The Financial News article 访谈 | 王景武:坚决打好防范化解重大金融风险攻坚战
5. US Senate panel investigates Confucius Institutes
Senate panel wants Chinese-funded institutes to change or leave U.S. | Reuters:
China has provided over $158 million to U.S. schools for Confucius Institutes to promote Chinese culture, U.S. Senate investigators said on Wednesday, releasing a report saying the centers have acted as tightly controlled propaganda arms for Beijing and should be changed - or shut down..
The Senate's Permanent Subcommittee on Investigations spent eight months investigating the Confucius Institutes, which were created in 2004 to promote Chinese language and culture at schools and universities around the world.
Here is the report - Majority and Minority Staff Report - China's Impact on the U.S. Education System
Video of the hearing today
Senate Panel Takes Aim at China-Funded Education Programs in U.S. - WSJ $$:
The eight-month Senate investigation drew from communication with more than 100 schools, including reviews of contracts for 15 U.S. schools, their Chinese partners and Hanban, as well as interviews with Confucius Institute officials.
It faults both schools and government agencies for not more carefully vetting Confucius Institutes and their instructors. "There is no other comparable arrangement with a foreign government in U.S. higher education," the report says...
It also said that while Education Department rules require colleges and universities to report foreign gifts of $250,000 or more, nearly 70% of U.S. schools that hit that threshold from Hanban didn't properly do so.
U.S. GAO - China: Agreements Establishing Confucius Institutes at U.S. Universities Are Similar, but Institute Operations Vary:
GAO reviewed 90 agreements and found they describe generally similar activities, funding, and management. For example, the institutes primarily receive funding from Hanban and the U.S. school, and do not receive direct U.S. federal funding. GAO also examined the agreements for language on application of school polices to the institutes, curriculum, and confidentiality, among other things. One-third of the agreements explicitly addressed how U.S. school policies apply to institutes, and a few addressed curriculum. Officials GAO interviewed at case study schools noted that U.S. school policies, including policies on matters such as curriculum, apply to institutes at their schools, though we found schools vary from one another in institute activities and use of resources, including teachers and teaching materials. While 42 of 90 agreements include language indicating that the document was confidential, some agreements were available online or are shared upon request. Some officials at schools that did not post agreements online said this was consistent with handling of other agreements.
6. Demographics
China has more people aged 60 and above than under 15 - Global Times:
The population aged 60 and above in China has for the first time surpassed those under 15, showing how the country's demographic structure is aging.
According to the statement released by National Bureau of Statistics (NBS) on Thursday, the population of China was about 1.4 billion at the end of 2018. Among them, the number of people aged 15 and under is 248.6 million; those who aged 60 and above are 249.5 million.
The NBS report - Statistical Communiqué of the People's Republic of China on the 2018 National Economic and Social Development
China's Employment Population Shrinks for First Time Ever - Caixin"
At the end of 2018, the number of people employed fell to 776 million, a drop of 540,000 from 2017, according to annual census data published Thursday by China's National Bureau of Statistics (NBS).
The working-age population, or people between the ages of 16 and 59, also shrank — for the seventh consecutive year, down a total of 2.8% from 2011 to 2018, a clear sign that China's population is aging rapidly.
7. China's massive Internet
China has 829 mln netizens, short-video users account for 78.2% - ECNS:
The number of Internet users in China hit 829 million at the end of December, according to a report from the China Internet Network Information Center (CNNIC).
As of December, the number of short video users reached 648 million, 78.2 percent of China's total Internet users.
The increase brought the country's Internet availability rate to 59.6 percent, with 26.7 percent of Internet users or 222 million living in rural areas, a year-on-year growth of 6.2 percent.
The cumulative mobile Internet traffic reached 71.1 billion GB, an increase of 189.1 percent. The average time people spent online reached 27.6 hours per week in last year...
Online shoppers numbered 610 million, up 14.4 percent, and about 592 million shopped with their mobile phones, an increase of 17.1 percent. Online payment users also rose by 13 percent year on year to reached 600 million.
According to the report, the number of Chinese Internet users using online financial services also increased by 17.5 percent to 151 million.
The report - 第43次《中国互联网络发展状况统计报告》
8. More on China's global governance ambitions
Mapping China's Global Governance Ambitions - Center for American Progress:
The United States and other democratic nations need to understand Beijing's vision for the global governance system and how that vision differs from the prevailing liberal democratic order, which recognizes limits to state authority, such as binding international law and unalienable individual rights. It is difficult to map China's true intentions based on Xi Jinping's international speeches, which make fuzzy promises that can be spun many different ways, such as his frequent claim that China aims to make the global governance system more democratic in order to "build a community of shared future for mankind."2 Yet when viewed through President Xi's domestic messaging and the actions Beijing has taken thus far to implement its global agenda, a clearer picture emerges. The current global governance system is rules-based, and it privileges liberal democratic values and standards; Beijing's alternative vision is a system based on authoritarian governance principles in which nations negotiate issues bilaterally instead of following common rules and standards. From a liberal democratic perspective, if Beijing succeeds in bringing about that vision, the world will be less free, less prosperous, and less safe.
This report aims to provide a deeper and more nuanced perspective on China's real global governance intentions by mapping what President Xi and other Chinese leaders are saying to their domestic audience and how Chinese foreign policy scholars interpret those statements. In the Chinese political system, internal leadership statements are particularly revealing because they convey Beijing's intentions to officials throughout the Chinese Communist Party and government hierarchy. This report will analyze that internal messaging alongside the empirical pattern of actions Beijing has taken thus far in order to assess the risks and opportunities that China's new activism presents for the current liberal order.
As I wrote yesterday about the 5th in a People's Daily series on "grasping the new implications of developing our nation's important strategic opportunities参与全球经济治理体系变革--观点--人民网"
I choose to translate 变革 as change or transformation, not reform. The CCP calls reform and opening as 改革开放 not 变革开放 so there seems to be a very important distinction between 改革 and 变革. There is still a debate about China's goals in shaping global governance systems, whether it wants to reform them to Beijing's advantage or transform them to Beijing's advantage. I think there is a difference, and from everything I have heard and read I believe transformation, not simply reform, is the goal.
A lot of this we have heard and read before, but it is worth revisiting given its placement and the current state of the world, and the hubbub in certain pundit circles that Xi is somehow chastened and pulling back on initiatives like BRI...
I am sure some of you disagree, so please argue with me in the comments
China February factory activity shrinks to three-year low, export orders worst in a decade | Reuters The official Purchasing Managers' Index (PMI) fell to 49.2 in February from 49.5 in January, pointing to a contraction in activity for the third straight month, according to data released by the National Bureau of Statistics (NBS) on Thursday. The 50-mark separates growth from contraction on a monthly basis. Analysts surveyed by Reuters had forecast the manufacturing gauge would stay unchanged from January's 49.5. China's factory activity has been generally softening since last May. // Comment: Bad data, but caution over-reading given the New Year Holiday
Prepare to Hold More Chinese Stocks as MSCI Decision Arrives - Bloomberg MSCI will say if it decided to lift a cap on free-float-adjusted market value to 20 percent from 5 percent for yuan-denominated stocks. The implementation would occur in two stages this year and mean that A shares made up 2.8 percent of the MSCI Emerging Markets Index by August, according to a consultation paper. MSCI will also decide whether to make equities listed on the tech-heavy ChiNext board eligible from May, and mid-cap stocks from 2020.
Chinese Stocks Are Suddenly the World's Best Trade in February - Bloomberg In the 15 days they've been open in February, China's financial markets have delivered spectacular returns for equity investors. The tech-heavy ChiNext stands out with a 25 percent surge, its best month on record, with all 100 members rising. The Shanghai Composite has added 14 percent, better than any other major equity index worldwide and its widest outperformance relative to global stocks since April 2015.
Chinese consumers: your country needs you | Financial Times $$ ...closer analysis suggests fears about falling consumption are exaggerated. The reasons behind the drop in car sales are often sector-specific factors, such as tax cuts being reversed. Likewise, retail sales offer an imperfect picture of household spending. The figures include corporate and government purchases, while as Chinese households become more affluent, more of their spending goes on services than goods. Mr Zhang, for instance, says 15 per cent of the family's household income is spent on tourism. Their daughter's education is another big expense
Chinese investors withdrawing from overseas property market - ECNS Chinese investors have been increasingly withdrawing from overseas real estate markets in recent years following tightened loan policies, said a report by global real-estate services company Cushman & Wakefield.
New tech board gets ready for launch - China Daily Dong Dengxin, director of the Finance and Securities Institute at the Wuhan University of Science and Technology, expects that both the detailed rules and the list of the first companies involved are likely to come out in March.
Starbucks' China rival Luckin Coffee taps three banks for U.S. IPO: sources | Reuters .
Yicai Global - Short Seller Citron Research Loses Suit, Is Banned From Hong Kong for Five Years
Xi's article on reform of national supervision system to be published - Xinhua In the fifth issue of the Qiushi Journal, to be published Friday 在新的起点上深化国家监察体制改革 and some highlights 习近平谈公权力监督 这些重点所有公职人员要牢记--专题报道-人民网
Xi highlights building of Marxist learning party, nurturing of love of learning - Xinhua President Xi Jinping has called for expediting the building of the Communist Party of China (CPC) into a Marxist learning party and the nurturing of a love of learning in the people. Xi, also general secretary of the CPC Central Committee and chairman of the Central Military Commission, made the remarks in a preface he wrote for a series of textbooks designed as learning and training materials for officials across the country, an official statement said Thursday. // 习近平:第五批全国干部学习培训教材序言
NPC Calendar: March 2019 – NPC Observer On March 4, the Session will convene for a preparatory meeting to select members of the Presidium (which will preside over the Session) and to finalize the Session's agenda. The Presidium will then immediately meet to decide on (among other matters) the Session's daily schedule. Shortly thereafter the Session is expected to hold its first press conference. We expect the NPC's 2019 annual session to close around March 15. On that last day, we expect it to approve the Foreign Investment Law.
China Focus: Key economic terms for upcoming "two sessions" - Xinhua Following is a list of key economic terms that will outline China's plan in maintaining stable 2019 growth: GDP growth; Fiscal deficit; Tax reduction; Monetary stance; Job creation; Poverty alleviation; Trade growth; Property market; Science and technology innovation board; Foreign investment
China's new veterans' law to be reviewed at National People's Congress next week | South China Morning Post Li Xiao, a 64-year-old veteran campaign leader, said he had been told that the draft, which comprises 83 articles in 11 charters, had been submitted to the National People's Congress (NPC), which begins its annual session on Tuesday. "This is the law the veterans have been waiting for for decades, with many retired senior military officers pushing for it since the 1970s. Now, we finally see a ray of hope," said the former artillery soldier who was involved in China's nuclear tests from 1978 to 1981.
王永康任黑龙江省委常委、省政府党组成员(图/简历)--组织人事-人民网 Xian Party Secretary Wang Yongkang moves to Heilongjiang to become a member of the provincial Party committee
Former Party official Li Rui passes away - Xinhua Li Rui, a former member of the Communist Party of China (CPC) Central Advisory Commission and former deputy chief of the Organization Department of the CPC Central Committee, died of illness in Beijing on Feb. 16 at the age of 102, said an official statement Thursday. // Xinhua- 原中顾委委员、中组部原副部长李锐在京逝世 享年102岁
Chinese law books written by judge behind stolen court papers scandal recalled by publisher | South China Morning Post Law books written by a Chinese judge were removed from shelves after he made a surprise televised confession about his part in a scandal at the country's highest court that he helped bring to light. More than 10 books written by Wang Linqing, a former assistant judge to the Supreme People's Court, were recalled by his publisher on Friday, employees at the company and bookstores said.
十三届全国人大一次会议专题--人民网 People's Daily site for the Two Meetings
Former executive of China Development bank drummed out of Party for severe discipline violations - Global Times Guo Lin, a former member of the China Development Bank's administrative committee, was found to have severely violated anti-corruption discipline during his stint as head of the bank's Tianjin branch, the CPC Central Commission for Discipline Inspection and the National Supervisory Commission said in a statement posted on its website.
Overseas returnee selling homegrown marijuana detained - Global Times After returning to China, Su, while smuggling marijuana from the US, started to learn how to grow marijuana from an English-language website, got marijuana seeds illegally and began growing the plant. According to Su's neighbors, Su always went in and out the apartment with peers and the apartment often smelled weird. Actually, Su lied to his parents twice to get more than 20,000 yuan ($2989) to buy marijuana seeds and growth equipment to harvest marijuana. Relying on Su's care around the clock, the survival rate of the marijuana was nearly 70 percent.
中共中央印发《中国共产党重大事项请示报告条例》--时政--人民网 Central committee issues "Regulations of the Communist Party of China on Asking for Instructions and Reporting Major Matters"
"学习强国"上海学习平台在PC端、手机客户端正式上线_浦江头条_澎湃新闻-The Paper
Inside the Investigation That Led to Prostitution Charges Against Robert Kraft - WSJ $$ Other clues about the wrongdoing included the women working at the spas only speaking dialects of Chinese. Masseuses are licensed in Florida after an exam that is only administered English and Spanish. Many of the licenses, authorities determined, were fraudulent. Subpoenas gave police access to bank accounts and other financial records of the spa operators. Mr. Snyder says the records showed upward of $20 million was flowing back and forth to parties in China.
China says difficulties in U.S-North Korea talks unavoidable | Reuters Difficulties in U.S.-North Korea talks are unavoidable, but China hopes both sides can continue their dialogue, the Chinese government's top diplomat told a visiting senior North Korean diplomat on Thursday, state media said.
Trump: China's been a 'big help' in US-North Korea engagement [Video] but notes that the hermit kingdom calls "its own shots."
Next stops: Europe and United States. Chinese President Xi Jinping to head to Italy, France and Florida on trade and tech missions | South China Morning Post After the annual National People's Congress gathering in Beijing, Xi is expected visit Italy in late March and then France, with another planned stop at US President Donald Trump's Mar-a-Lago resort in Florida.
Donald Trump, human rights activist? Meet the Chinese dissidents sticking up for the US president | South China Morning Post
Jaw-Jaw: China's Great Power Disease - War on the Rocks Podcast - Edward Luttwak and Brad Carson
China to hold naval parade on April 23 in Qingdao - CGTN to celebrate the 70th anniversary of the founding of Chinese People's Liberation Army Navy.
App launched to promote preferential policies across Taiwan Strait - Xinhua Users can check on the implementation of 31 preferential policies for Taiwan in the mainland's 25 provincial-level regions on the app. The app also has a Q & A section. The app, released by the Cross-Straits Internet Exchange Committee of the Internet Society of China, will soon be available in app stores.
'Bohemian Rhapsody' Gets China Release | Hollywood Reporter Sources also tell THR that at least one minute of cuts will be made to the film, removing portrayals of drug use and several intimate kisses between Malek's Mercury and other male characters.
KKR, China's Tencent eyeing bids for Universal Music - sources | Reuters Universal will generate roughly 1.5 billion euros of free cash flow excluding interest payments in 2023, Deutsche Bank forecast. Tencent Music Entertainment Group, a subsidiary of China's biggest gaming and social media firm Tencent Holdings Ltd, has an existing licensing agreement with Universal and wants to strengthen its collaboration with a partial acquisition, the sources said, cautioning that no deal was certain.
Login - TikTok predecessor gets record FTC fine TikTok, a short-form video app owned by Chinese tech giant Bytedance, has agreed to a $5.7 million settlement with the U.S. Federal Trade Commission for illegally collecting personal data from children, Axios' Sara Fischer reports...It's the largest settlement from a violation of the Children's Online Privacy Protection Act (COPPA) in the law's 20+ year history.
Chinese 'Deepfake' Creator Says Videos Meant to Educate Public - Sixth Tone "My initial intention was to help more people hear about this technology and protect celebrities from videos of forged, negative content," the videos' creator, who says his surname is Xiao wrote Wednesday on video-sharing platform Bilibili. "The [videos'] virality this time should be a warning to us all." In two videos that have gone viral since they were released on Bilibili in mid-February, Xiao digitally grafted mainland actress Yang Mi's face onto Hong Kong star Athena Chu's body.
'Farewell, so long': video of Taiwanese TV host's assisted suicide fires up euthanasia debate | South China Morning Post A video of Taiwanese TV host Fu Da-ren's death by assisted suicide at a clinic in Switzerland has gone viral, raising the profile of the debate on euthanasia in China and questions about dignity at life's end...In the video, which became public on Taiwanese media this week, Fu, surrounded by his family, is seen holding a cup of medicine. A volunteer with Dignitas, an organisation that enables assisted suicides, tells him to take the drug as quickly as he can because it has a bitter taste.
Coal Consumption Increases Despite Drop in Share of Energy Mix - Caixin China's overall coal consumption and carbon dioxide emissions climbed last year, even as renewable energy generation made gains and coal's share of the nation's energy mix fell below 60% for the first time. Coal's share of the overall energy mix fell to 59%, but China consumed 1% more coal year-on-year, according to data releasedby the National Bureau of Statistics today. Solar and wind made big advances, with total installed capacity increasing by 33.9% and 12.4% respectively. // 2018年国民经济和社会发展统计公报
Ministry announces new measures to clean up Qomolangma - ECNS The measures included strengthening daily inspection of the Qomolangma nature reserve, regulating the environment at tourist attractions in areas below the elevation of 5,200 meters and also a task force to regularly clean up trash left by visitors. Other measures include offering new garbage bins, building environmentally friendly toilets and providing portable toilets for each mountaineering team, said Liu.
Tomb-raiding themed drama drives pangolin purchase - Global Times According to a recent documentary, Searching for the Last Pangolin, published on World Pangolin Day on February 16, pangolin products were found to be the new favorites for tomb-raiding themed drama followers. Pangolin claws are crafted into an ornament called "mojinfu." Some believe wearing mojinfu can exorcise evil spirits as depicted in the fantasy novel Guichuideng, also known as Ghost Blows Out the Light, depicts.
International Math Competition Defeat Prompts Soul Searching in China - Caixin China placed a mediocre sixth at the 2019 Romanian Master of Mathematics (RMM), a major math competition for pre-university students. The U.S. won the championship for best team, while the highest individual prize went to an Israeli candidate. Math competitions like the RMM are serious business in China, where participation can give students a leg up in university admissions. China's defeat on Monday prompted social media users to ask if recent Ministry of Education curbs on math competitions were misguided.


If you were to believe the spin in the New York Times the other day, you would think that Marc Tucker, who has long been Hillary Clinton's main man on education issues, had decided that high stakes testing was a thing of the past: "Tucker would not abolish tests, but he would have fewer of them. And they would have a different purpose."
But here is the real bottom line message for the new Tuckerism, which looks remarkably like the old Tuckerism.


Roy Fowler, Study for Thallos (2013). Courtesy of Fort Gansevoort.
Along the Embarcadero, overlooking the bay, the UNTITLED San Francisco fair has settled into its third home in as many years: Pier 35, a cruise ship terminal with a decidedly industrial chic vibe. No fewer than 55 dealers from 16 cities have made the trip, hoping to lure in wealthy tech elite and other local collectors.
Thursday's opening followed on the heels of the gala opening at FOG Design+Art, now its sixth year. The two fairs "wanted to work together to create more of an art-fair week for San Francisco," UNTITLED's director Manuela Mozo told artnet News.
Dealers are eager to break into the San Francisco market. At least one gallery, Espai Tactel of Valencia, Spain, is even making its first stateside appearance. "We think San Francisco is interesting; it is a multicultural town," said the gallery's co-founder Juanma Menero.
Another first-timer, Blain|Southern of London and Berlin, is doubling down on San Francisco with booths at both of the city's fairs. At UNTITLED, they are near the entrance, but have hidden Mat Collishaw's piece All Things Fall behind a wall in a darkened chamber, where a strobe light transforms the spinning work into a sculptural zoetrope.
Visitors who spotted the hidden room were instantly transfixed by the optical illusion of animation, based on Peter Paul Rubens's The Massacre of the Innocents. The 3D-printed work has been shown at the Galleria Borghese in Rome, and is priced at £350,000 ($451,000).
"We wanted to focus on bringing him to the US market," gallery sales consultant Ashley White told artnet News, noting that the first of the two editions of the work belongs to artist Tony Cragg, and that Damien Hirst owns a similar piece by the artist.
Roula Partheniou, Bubble Formation (2018). Photo courtesy of MKG127.
While there were no takers yet on Collishaw, a number of galleries reported early sales during the fair's Thursday preview. Many were photographs, which has long been a medium of choice for major San Francisco collectors, and other works below $20,000.
Toronto's MKG127 quickly sold two of Roula Partheniou's painted wooden sculptures, which realistically ape colorful objects such as raffle tickets and boxes of candy, for $5,500 and $2,800.
By the end of day one, New York's Fort Gansevoort had sold five of nine paintings from its solo presentation Roy Fowler, a former California surfer kid who makes striking blue and white works depicting crashing ocean waves. At the fair, his canvases are priced between $10,000 and $20,000.
Five Car Garage from Santa Monica dedicated its booth to the romanticism-tinged paintings of Alison Blickle, and sold two canvases for between $8,000 and $10,000.
Christian Schumann, Biota (2016). Photo courtesy of Neuman Wolfson Art.
"UNTITLED kind of curates your booth and tells you the artists you think will do well," Belinda Neuman, the co-owner of first-time exhibitor Neuman Wolfson Art of New York, told artnet News. The gallery presented a two-man booth of Michael Bevilacqua and Christian Schumann, whose star-covered panel painting sold for $32,000.
"Christian went to art school in San Francisco and Michael grew up in California and is in the collection of SFMOMA, so they seemed like a natural choice here, because it's a regional fair," Neuman added. The gallery also brought works on paper starting at $3,000 in an effort to accommodate a range of budgets.
Masako Miki's booth with the Berkeley Art Museum Pacific Film Archive at UNTITLED, San Francisco. Photo by Sarah Cascone.
Even lower on the price spectrum, the Berkeley Art Museum Pacific Film Archive was doing brisk business in their eye-popping booth selling $100 prints by Masako Miki. Her exhibition "Masako Miki/MATRIX 273" is on view at the museum through April 28, and the artist is leading a poster-making workshop at the fair this afternoon to help guests create signage for tomorrow's Women's March.
Trends can be hard to spot at art fairs, but there were a number of booths highlighting photography. "The history of collecting in San Francisco is traditionally more geared toward photography," noted the Seattle-based dealer Mariane Ibrahim, who was showing exclusively photos by Ayana V. Jackson.
Ayana V. Jackson, Sleep to Dream (2016). Photo courtesy of Mariane Ibrahim Gallery.
The striking presentation featured toile wallpaper on the booth walls, contrasting an idyllic county scene with photographs referencing histories of slavery and the objectification of the black female body, as well as contemporary concerns such as Black Lives Matter. The works are priced between $3,800 and $12,000.
Meanwhile, rising star Devan Shimoyama had an entire wall at New York's De Buck Gallery, with four photographs from the artist's residency in New York's Fire Island, also currently on view at his solo show at the Andy Warhol Museum in Pittsburgh.
Devan Shimoyama, Birth of the Sunrise Summoner (2015). Photo courtesy of De Buck Gallery.
"Devan's really taken off the last year and a half," said owner David De Buck. "People seem to be very attracted to this work." The gallery also sold a photographic collage by Rashaad Newsome for between $15,000 and $20,000.
Paris's onestar dedicated its entire stand to Daniel Gordon's series "Au Bon Marché." The photographs capture still-life arrangements the artist created by collaging found photographs of everyday objects into 3D versions of themselves, instantly recognizable but delightfully distorted. Each print cost €2,000 ($2,270) for the first edition and €2,500 ($2,840) for the second edition.
Daniel Gordon, Artichoke, from the series "Au Bon Marché." Photo courtesy of onestar.
Gordon originally discarded the sculptures after creating the photographs, but the originals are now for sale as well. "We said, 'stop destroying it! People should see these objects themselves,'" said Darlene Lin, onestar's director of sales.
That's not the case with Jasper de Beijer, whose photographs are the only record of his carefully constructed architectural models, which are about two-thirds the size of the final, larger-than-life images.
Jasper de Beijer, 12-26-2004 (from Mr. Knight's World Band Receiver) (2014). Photo courtesy of Asya Geisberg Gallery.
New York's Asya Geisberg Gallery had already sold one of his works for $10,000. "Jasper is actually the only photographer we have in our program at the moment," said gallery manager Hanna Fruchtenicht, citing the medium's popularity among the local collecting community.
Lamarche-Ovize, Elisée (2), 2018. Photo courtesy of Galerie Lefebvre et Fils.
Despite a great deal of optimism among dealers at both fairs, not everyone is convinced that San Francisco art fairs can be money-makers for galleries. One Bay Area dealer who has shown with UNTITLED in Miami Beach—reliably selling out the gallery's booth—but has yet to commit to showing in their hometown, told artnet News that they weren't sure they could sell enough in San Francisco to cover the cost of the booth.
But if the market hasn't totally caught fire, it is certainly on its way.
UNTITLED, San Francisco, is on view at Pier 35, 1454 the Embarcadero, San Francisco, January 17–20, 2019.


text:
Photography by Ashley Thompson for Our Era at Blonde Studios // Design by Yoora Kim//Instagram
"Our Era" x Instagram Creators Talk Mental Health
Gen-Z Founded Magazine "Our Era" and Instagram Creators Bring Awareness to Intentionality Online
Gen-Z magazine Our Era and Instagram teamed up to produce four cover stories featuring emerging creators who are using social media to bring awareness to mental health.
By Samantha Olson Published: May 13, 2022
What seemed to be another gloomy Tuesday in NoMad, New York City turned out to be an eventful day full of love and light at Blonde Studios where a team made up of a hairstylist, makeup artist, wardrobe stylists, editors, creative directors, videographers, and creators joined forces to put together a colorful shoot that was just as vivid and complex as the individuals behind Our Era. During the COVID-19 pandemic, Lucy Ivey decided that young people deserved a creative space where they could gain valuable experience in writing, editing, photography, and artistic collaboration. She was just 15 years old when she birthed, her own magazine, Our Era. The publication has taken to new heights since it was founded two years ago.
"I turned to a lot of people I knew at the time when I was like 14. I founded a nonprofit called Teens Speak Up and it helped make sure teenagers could speak about issues they're passionate about. We started a podcast and then it led to a site where people could do op-eds and everything," Lucy explained to Seventeen. "That kind of started my passion for editorial writing, and then there were a lot of creative people on that team and I told them about this idea I had for Our Era and some of them came along."
Our Era's 18-year-old features editor, Ella Edwards, also explained that people reach out to their team in hopes of joining — including herself when she made the move to the east coast from Los Angeles. "I personally reached out because when I moved to New York and I started college, I really missed working for a magazine," Ella tells Seventeen.
Eighteen-year-old Lucy Ivey is the founder and editor-in-chief of Our Era, a magazine catered to fashion, politics, music, social justice, mental health, and identity.
Ashley Thompson for Our Era//Instagram
Now 18, Lucy has spearheaded collaborations with brands like Steve Madden for Our Era, which has featured an impressive lineup of cover stars like TikTok sensation Emmy Hartman, High School Musical: The Musical: The Series star Dara Reneé, and indie musician Claud. Next up on their features roster is a foursome of Instagram creators who use their platforms to promote positivity and encourage their followers to prioritize their mental health. The Gen-Z magazine has joined forces with queer Asian-American artist Kim Saira, artist and writer Tauri Janée, spiritual creator Mikah Jones, and queer indigenous Inuk Serqininguak Ketura to have open conversations about intentionality online, or the purpose behind their posts and how it affects their mental health.
"Intentionality to me means making sure that you're consuming content that is good for you and taking care of yourself on the internet in the way that if you're following someone who doesn't bring you joy or makes you compare yourself... it's important to maybe mute those people and be mindful about how much time you're spending on social media," Lucy explains.
Ella adds that the team at Our Era feels they should be providing Gen-Z with effective resources that cater specifically to them. "We're creating content for a generation that spends so much time on their phone, so we have a responsibility to those people to make sure they're consuming content that services them, helps them, and uplifts them. This collaboration with Instagram is really important because it's coming from the people that they're trying to cater towards."
As for the creators featured on the covers of Our Era's special issue in collaboration with Instagram, Lucy recalls the process of seeking them out and why they were the perfect fit. "These four creators really stuck with us because of how they create their content and their interest in art. Their takes on mental health were really, really interesting," she says.
Kim Saira echoes that sentiment, sharing that she was drawn to the project because of each creator's different point of view when it comes to mental health and how that journey looks different for Black, brown, and queer folks. "I think that whatever healing means to someone might not mean the same to someone else. So, I really love the fact that Our Era and Instagram are shining a light on Black and brown voices and what our wellness looks like," Kim tells Seventeen. "I really appreciate having that amplification to younger audiences because it's something that I feel like I definitely would've liked when I was a lot younger."
Each featured creator has their own lane when it comes to their content — for instance, Mikah encourages wellness through live guided meditations while Serqininguak shares a lot about purpose and what that means for indigenous Peoples. Meanwhile, Kim and Tauri turn to art forms like graphic design, writing, and dance to spread awareness about mental health and prioritizing wellness.
The Our Era x Instagram cover stars Serqininguak Ketura, Tauri Janée, Kim Saira, and Mikah Jones spoke about their experiences with social media and mental health in the special issue.
"[Content creation] has encouraged me to create distance between myself and social media. I value my well-being and I realize that creating space between myself and the virtual world serves me well," Tauri explains. "Of course, I still value the relationships and the opportunities that have presented themselves and the connections that form on these platforms. But I've learned that those things are equally as accessible offline and that there are other ways for me to feel affirmed."
A 2019 survey by the American Psychological Association found that Gen-Z is more likely to report they have fair or poor mental health. It also revealed that Gen-Z is more likely to report they have received treatment from a mental health professional or gone to therapy. Ashley Thompson, the 19-year-old photographer behind the Our Era x Instagram cover shoot, credits Gen-Z's continuous transparency on social media for encouraging individuals to seek help. "Obviously, mental health has been an issue for however long we've been alive but I think that our generation has been the most indulged in social media," Ashley tells Seventeen. "Everyone has started sharing so much and having all these platforms to share on that they've been able to talk about where they're at mentally." She also explains that the rise of social media platforms has helped people speak out and create a domino effect where a community is formed and others feel comfortable in their own transparency.
"I think that knowing that there are people who can resonate with what you're going through is very reassuring. It's so important to ask for help because you can't handle [certain struggles] on your own. Like, you can tell yourself you can, but you really can't," Lucy adds.
As for navigating our own mental health journies, Ella suggests that progress isn't linear and that as complex individuals, we're allowed to feel certain emotions. "You need to allow yourself to be angry and you need to allow yourself to make mistakes. Above all else, allow yourself to change. If you're the same person that you were a year ago, then nothing happened — no change was made," she says. "You aren't supposed to know everything right away and you should allow yourself to have those dips because you're always gonna come out the other side and learn something from it."
Our Era's special issue with Instagram about intentionality online is available now. Read all four cover stories at www.oureramag.com.
Studio: Blonde Studios
Photographer: Ashley Thompson
Our Era Graphic Designer: Charlie Sue Birznieks
Stylist: Bernadet Davood with assistance from Tatiana Dominguez
Producer: Lucy Ivey
Production assistant: Ella Edwards
Hair: Mariah Molina
Makeup: Danielle Parkes
On-set assistant: Hannah-Mia Hinds
Samantha Olson
Sam is an assistant editor at Seventeen, covering pop culture, celebrity news, health, and beauty. When she isn't draping her cheeks in blush, you can probably find her live-tweeting awards shows or making SwiftToks.
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redpajama_set_name: RedPajamaCommonCrawl


Brainchild of Ayaz Weldon and Anis Weldon, it's an exciting concept everyone's been talking about in the Indian beauty circuit. Very soon Mumbai will get the best of beauty experts under one roof!
Origin of TABS : Tabs mean small pieces!! We are aiming at getting the pieces (Tabs) together under a single roof.
TABS is more of a platform for entrepreneurs in the cosmetics/beauty industry who has the zeal to learn or who already has knowledge and expertise but does not have resources to accomplish it.
Anybody who enter the premise for any service will get it. Be it Hair, Skin, Nails, Make up , Make over, Personal Grooming, Mehendi and so on.
Anybody who wants to learn will get the connection with an industrial expert in that field.
We have diverse trainers in terms of professional level, expertise, knowledge associated with us in the diverse fields of Hair, Skin, Nails and Make up.
We have training classes with short courses on topics of Hair styles, Hair do's, Skin machineries, Nail art, Make up – looks, and product based training for people also who are working but can benefit their customers with the proper use of the products as well.
We have associated with Rashida Kapadia who is an Image consultant to conduct her "A new year a new you" personal grooming for New Years eve happening on December 20th 2015.
Similarly we will tie up with experts and have a vision of connecting the people with a Solution that they are looking for.
Do you have Expertise but do not have resources to start your own Salon Business?
Do you go through the hassle of calling your clients to different locations for services while freelancing?
Do you need a flexibility to work at your own time and days you feel like working?
Do you want a platform to show your skills and make your own business with the help of the professional?
Do you want to have the ownership of your own space with not having to worry about the bills, the stock and the maintenance.
Start your own Salon Business with us with no initial investment, interiors, stocks, bills or maintenance, In short a hassle free way of making your own clients at your own convenience.
a) Chair, Mirror, Electrical, equipment's, accessories, Consumables (in-house). b) Interiors, Maintenance, Advertisements, business Consulting, Cross-Services (Hair, Skin, Nails, Make-up). c) In – house mini store for all your consumables with a Owners-special Retail Margin tailored for your Business.
TABS presents various grooming services for our customers under one roof. We have a platform where we have variety to offer you not only in terms of different hair do's, nail art styles, facials or make over but also variety in the number of stylists to chose from.


Response: Heart failure is a global epidemic affecting greater than 35 million individuals worldwide. Readers should take away from this report the fact that once an elderly patient is hospitalized with a diagnosis of heart failure, their long-term risk of death and re-hospitalization is significantly greater than the general population. Moreover, these findings apply across the spectrum of ejection fraction. This finding is important to the public because it underscores the serious nature of the diagnosis of heart failure and the long-term risk associated with it.
Response: The next steps are to determine the specific causes of death for these different subgroups and then determine other potential treatment strategies to improve their outcomes.
Response: This investigation is the result of a collaborative effort from Get with The Guidelines-Heart Failure Investigators and the Duke Clinical Research Institute (DCRI). No financial disclosures for myself.
This entry was posted in Author Interviews, Geriatrics, Heart Disease, JACC, UCLA and tagged AHA16, ejection fraction, heart failure by Marie Benz MD FAAD. Bookmark the permalink.


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Viral titer (plaque assay)
Cytokine assay
Northern (RNA) blot analysis
Cytotoxicity of ribavirin
Effect of ribavirin on the growth of MHV-3 in macrophages
Effects of ribavirin on functional PCA and transcription of mRNA for fgl-2 in MHV-3-infected macrophages
Effect of ribavirin on production and mRNA transcription of TNF in macrophages
Effect of ribavirin on production and mRNA transcription of IL-1 in macrophages
Effect of ribavirin on Th1/Th2 cytokine profiles
Ribavirin Inhibits Viral-Induced Macrophage Production of TNF, IL-1, the Procoagulant fgl2 Prothrombinase and Preserves Th1 Cytokine Production But Inhibits Th2 Cytokine Response1
Qin Ning;
Qin Ning
*Medicine,
Multiorgan Transplant Program and Departments of
Deron Brown;
Deron Brown
Jean Parodo;
Jean Parodo
†Surgery, and
Mark Cattral;
Mark Cattral
Reginald Gorczynski;
Reginald Gorczynski
Edward Cole;
Edward Cole
Laisum Fung;
Laisum Fung
Jin Wen Ding;
Jin Wen Ding
Ming Feng Liu;
Ming Feng Liu
Ori Rotstein;
Ori Rotstein
M. James Phillips;
M. James Phillips
‡Pathology, The Toronto Hospital and The University of Toronto, Toronto, Canada M5G 2C4
Gary Levy
Address correspondence and reprint requests to Dr. Gary Levy, 621 University Avenue, 10th Floor, Room 151, Toronto, Ontario, M5G 2C4, E-mail address: fgl2@msn.com
Qin Ning, Deron Brown, Jean Parodo, Mark Cattral, Reginald Gorczynski, Edward Cole, Laisum Fung, Jin Wen Ding, Ming Feng Liu, Ori Rotstein, M. James Phillips, Gary Levy; Ribavirin Inhibits Viral-Induced Macrophage Production of TNF, IL-1, the Procoagulant fgl2 Prothrombinase and Preserves Th1 Cytokine Production But Inhibits Th2 Cytokine Response1. J Immunol 1 April 1998; 160 (7): 3487–3493. https://doi.org/10.4049/jimmunol.160.7.3487
Ribavirin, a synthetic guanosine analogue, possesses a broad spectrum of activity against DNA and RNA viruses. It has been previously shown to attenuate the course of fulminant hepatitis in mice produced by murine hepatitis virus strain 3. We therefore studied the effects of ribavirin on murine hepatitis virus strain 3 replication, macrophage production of proinflammatory mediators including TNF, IL-1, and the procoagulant activity (PCA), fgl2 prothrombinase; and Th1/Th2 cytokine production. Although ribavirin had inhibitory effects on viral replication (<1 log), even at high concentrations complete eradication of the virus was not seen. In contrast, at physiologic concentrations (up to 500 μg/ml), ribavirin markedly reduced viral-induced parameters of macrophage activation. With ribavirin treatment, the concentrations of PCA, TNF-α and IL-1β all decreased to basal concentrations: PCA from 941 ± 80 to 34 ± 11 mU/106 cells; TNF-α from 10.73 ± 2.15 to 2.74 ± 0.93 ng/ml; and IL-1β from 155.91 ± 22.62 to 5.74 ± 0.70 pg/ml. The inhibitory effects of ribavirin were at the level of gene transcription as evidenced by Northern analysis. Both in vitro and in vivo, ribavirin inhibited the production of IL-4 by Th2 cells, whereas it did not diminish the production of IFN-γ in Th1 cells. In contrast, ribavirin had no inhibitory effect on TNF-α and IL-1β production in LPS-stimulated macrophages. These results suggest that the beneficial effects of ribavirin are mediated by inhibition of induction of macrophage proinflammatory cytokines and Th2 cytokines while preserving Th1 cytokines.
Ribavirin (1-β-d-ribofuranosyl-1,2,4-triazole-3-carboxamide) is a synthetic guanosine analogue that possesses a broad spectrum of activity against DNA and RNA viruses (1). Earlier studies have shown that ribavirin has immunosuppressive activity, reducing lymphocyte proliferation to mitogens in vitro and prolonging skin allograft survival in rats (2, 3).
Previous reports have shown that ribavirin attenuates the course of an experimental model of fulminant hepatitis in mice produced by murine hepatitis virus strain 3 (MHV-3)3 (4). Studies in our laboratory as well as others have demonstrated a causal relationship between macrophage activation, production of proinflammatory mediators such as TNF and IL-1 as well as the unique procoagulant activity fgl2 prothrombinase, and the subsequent development and pathogenesis of MHV-3-induced liver disease (5, 6).
To examine the mechanisms of the beneficial effect of ribavirin in this experimental model of viral hepatitis, we studied its effects on MHV-3 replication; its effects on macrophage production of TNF, IL-1, and PCA and Th1/Th2 cytokine production in vitro.
Female BALB/cJ, A/J, and C3H/HeJ mice, 6 to 8 wk of age, were purchased from Charles River Laboratories (St. Constant, Quebec, Canada). The animals were kept in microisolator cages and housed in the animal facilities at the Toronto Hospital, and were fed with standard laboratory chow diet and water ad libitum.
MHV-3 was obtained from American Type Culture Collection (Rockville, MD) and was plaque purified on monolayers of DBT cells and grown to titers of 2 × 107 plaque-forming units (PFU)/ml in 17 CL1 cells. Virus was harvested by centrifugation at 4500 × g for 1 h at 4°C and was assayed on monolayers of L2 cells in a standard plaque assay (5).
Peritoneal macrophages were harvested from BALB/cJ mice 4 days after i.p. administration of 1.5 ml of 3% thioglycolate (Difco Laboratories, Detroit, MI) as previously described (6). Macrophages were resuspended in RPMI 1640 (ICN Biomedicals, Costa Mesa, CA) supplemented with 2 mM glutamine (Sigma Chemical, St. Louis, MO) and 2% heat-inactivated FCS (Flow Laboratories, Mississauga, Ontario, Canada). Macrophages were >95% pure as determined by morphology and nonspecific esterase stain (7). Viability exceeded 95% by trypan blue exclusion.
A Th1/Th2 cell line was derived from C3H/HeJ mice that had been immunized with 1 × 108 irradiated (2000 rad) B10BR spleen cells and poly(IC) i.p. T cells were recovered from spleen, diluted, and incubated with 2 × 105 irradiated B10BR cells and IL-2 in 96-well U-bottom plates. Cells were fed weekly by adding fresh medium and rechallenged every 10 days with freshly harvested and irradiated B10BR spleen cells. Wells showing proliferation were transferred to T-25 tissue culture flasks. Stable T cells were then passaged and redistributed at limiting dilution under the same conditions (with fresh medium and irradiated B10BR spleen cells).
CD4+ Th1 (3E9.1) and Th2 (3B6.8) cell lines were derived from MHV-3-resistant A/J and MHV-3-susceptible BALB/cJ mice, respectively, as previously described (8). The cell lines were recovered from draining lymph nodes of mice that had been injected in the footpad with MHV-3. The cell lines were subtyped by flow cytometry and cytokine production assayed by the ability of supernatants to support proliferation of CTLL-2 and CT4.S cells in the presence of anti-IL-2 and anti-IL-4 mAbs, respectively. IFN-γ activity was determined by inhibition of plaque formation induced by MHV-3 on L2 cells (8). Stable T cells were passaged with fresh medium and irradiated splenic mononuclear cells as described previously (8).
The effect of ribavirin on viral titer were determined in a standard plaque assay (5). Briefly, monolayers of peritoneal macrophages from BALB/cJ were pretreated with either ribavirin (0–500 μg/ml) or medium (RPMI 1640), and 2 × 106 PFU of MHV-3 was then added. Following a 10-h incubation at 37°C with 5% CO2, cells were harvested, subjected to one cycle of freeze-thawing, and assayed for viral titers on monolayers of L2 cells.
MHV-3-infected macrophages, at a multiplicity of infection (MOI) of 2.5, were incubated with ribavirin for 8 h in supplemented RPMI 1640. Uninfected macrophages and MHV-3-infected macrophages without drug treatment were set up as negative and positive controls, respectively. Macrophages were evaluated for functional PCA in a one-stage clotting assay as previously described (5). Following incubation, samples to be assayed for PCA were washed three times with unsupplemented RPMI 1640 and resuspended at a concentration of 106/ml. Samples were assayed for the ability to shorten the spontaneous clotting time of normal citrated human platelet-poor plasma. Milliunits of PCA were assigned by reference to a standard curve generated with serial log dilutions of a standard rabbit brain thromboplastin (Dade Division, American Hospital Supply, Miami, FL). Media and reagents were assayed showing no PCA activity.
TNF.
MHV-3 (at a MOI of 2.5) or LPS (10 μg/ml, Sigma)-stimulated BALB/cJ macrophages were incubated with ribavirin (0–500 μg/ml) for 8 or 4 h, respectively. Supernatants were collected, and TNF-α concentrations were assayed by ELISA. Monoclonal hamster anti-murine TNF-α Ab (Genzyme, Boston, MA) was coated to ELISA plates overnight at 4°C. After being washed with Tris buffer (pH 8.0), plates were blocked with 100 μl of 6% BSA in each well for 1 h at room temperature. Following washing, 100 μl of standards or samples were added and incubated at room temperature for 3 h. Subsequently, 100 μl of polyclonal rabbit anti-mouse TNF-α Ab (Genzyme, IP-400) were added to each well, and plates were incubated at 4°C overnight. Goat anti-rabbit IgG alkaline phosphates (100 μl) (Jackson ImmunoResearch Laboratories, West Grove, PA) was added, and plates were incubated at room temperature for 1 h. Following washing, 100 μl of FSAP (diflusinal phosphate, 1/10 in substrate buffer) were added, and plates were incubated for an additional 10 min at room temperature with shaking. Plates were read by Cyber Fluor (Beckman Instruments, Fullerton, CA). Units were assayed by comparison to a mouse TNF-α standard (Genzyme).
IL-1.
MHV-3 (at a MOI of 2.5)- or LPS (10 μg/ml)-stimulated macrophages were incubated with ribavirin (0–500 μg/ml) for 8 or 4 h, respectively. Production of IL-1β in supernatants was determined by ELISA (R&D Systems, Minneapolis, MN). An affinity-purified polyclonal Ab specific for mouse IL-1β was precoated onto the microtiter plate. Standards, controls, and samples were added to the wells for 2 h at room temperature. After washing, an enzyme-linked polyclonal Ab specific for mouse IL-1β was added to the wells, and plates were incubated at room temperature for 2 h. Following washing, a substrate solution was added to the wells. After 30 min of incubation at room temperature, the stop solution was added, and plates were read by Titertek Multiskan MCC/340 Mk. Ribavirin was added to the supernatants from MHV-3- or LPS-infected macrophages at a final concentrations of 1–500 μg/ml before assay, no effects were found on the ELISA assay for TNF-α and IL-1β.
IL-4 and IFN-γ.
Production of IL-4 and IFN-γ in the T cell lines (9) was measured by bioassay. For the Th1/Th2 cell line, 2 × 104 cells were first stimulated with peritoneal cells from B10BR (C3H) and cultured with ribavirin (0–500 μg/ml) for 24 h in a final volume of 200 μl in RPMI 1640 supplemented with 10% FCS, 2 mM glutamine, 100 U of penicillin, and 100 μg of streptomycin (Flow Laboratories) per ml in 96-well flat-bottom microtest plates (Falcon Laboratories, Grand Island, NY). For the MHV-3-specific Th1 (3E9.1) and Th2 (4B6.8) cell lines, the lines were cocultured in growth factor-free medium with MHV-3-infected, irradiated splenic mononuclear cells as APCs, and the supernatants were collected 48 h later. Supernatants were collected for measurement of IFN-γ and IL-4 production by the ability of supernatants to inhibit or support the proliferation of Wehi 279 and CT4.S cells, respectively. Cultures were incubated for 40 h in a humidified CO2 atmosphere at 37°C, pulsed with 1 μCi of [3H]TdR (sp. act. 2 Ci/mmol; Amersham, Arlington Heights, IL), and harvested 26 h later onto glass fiber filters. Total cell-associated radioactivity was measured in a Beckman scintillation counter, and bioassay data were expressed as nanograms per milliliter for IFN-γ and units per ml for IL-4, derived from a standard curve with mouse recombinant IFN-γ and IL-4 (Genzyme). Ribavirin was added to Wehi 279 and CT4.S cells at different concentrations (1–500 μg/ml), showing no effect on the assay system (data not shown).
The effect of ribavirin on mRNA levels of the MHV-3-induced PCA, fgl2 prothrombinase, TNF, and IL-1 were assayed by Northern blot analysis. Macrophages (1 × 107) were pelleted in 1.5-ml Eppendorf tubes, and total cellular RNA was isolated by 8 M acid-guanidium hydrochloride extraction in a modified procedure described by Arnstein and Cox (10). RNA was resolved on an 1% agarose gel containing formaldehyde and transferred onto a nitrocellulose membrane (Bio-Rad, Oakville, Ontario, Canada). A 1.3-kb fgl2 prothrombinase cDNA (11), a 1.4-kb TNF cDNA (12), and a 2-kb IL-1 cDNA (13) were excised from plasmids, purified by agarose gel electrophoresis, and labeled by using a random-priming DNA labeling system (Pharmacia, Montreal, Quebec, Canada) with [α-32P]dCTP (sp. act. 3000 Ci/mmol; Amersham). Membranes were prehybridized for 5 h at 42°C in a mixture containing formamide (50%), 5 × Denhardt's solution, 0.2% SDS, 100 μg of denatured salmon sperm DNA per ml, and 5× SSPE. Hybridization was conducted overnight at 42°C in the same fresh mixture. The membranes were then washed under medium to high stringency conditions, and the membranes were exposed to Kodak XAR-5 film with intensifying screens for 16 to 24 h at −70°C. To confirm that equivalent amounts of RNA were loaded in all lanes, the membranes were probed with α-tubulin cDNA.
Statistical analysis was conducted by Student t test, and a p value of 0.05 or less was considered to be statistically significant. Results were reported as the mean ± SD for at least three separate experiments, each performed in triplicate.
Addition of ribavirin up to 500 μg/ml to peritoneal macrophages freshly isolated from BALB/cJ mice had no toxic effects as demonstrated by trypan blue exclusion (>95% viable).
The addition of ribavirin to macrophages from BALB/cJ mice inhibited MHV-3 replication as demonstrated in Figure 1. The inhibitory effects showed a dose-dependent pattern over the range 10 to 500 μg/ml. However, in macrophages, the maximal inhibitory effect of ribavirin was only 1 log, from 2.40 ± 0.02 × 107 PFU/ml to 1.55 ± 0.07 × 106 PFU/ml at a concentration of 100 μg/ml with no further increase at a concentration as high as 500 μg/ml compared with no ribavirin treatment. The inhibition was not due to toxic effects of ribavirin on cells as demonstrated by trypan blue exclusion.
Inhibitory effect of ribavirin on MHV-3 replication in BALB/cJ macrophages and L2 cells. Confluent monolayers of macrophages in six-well plates were infected with 2 × 106 MHV-3 in the presence or absence of indicated concentration of ribavirin for 10 h. For the pretreatment group, cells were incubated with ribavirin for 2 h before MHV-3 infection. Viral titers were measured on monolayers of L2 cells in a standard plaque assay. The bars represent the mean ± SD of four independent experiments compared with control (ribavirin, 0 μg/ml) by an unpaired t test (* two-tailed p < 0.01).
To determine whether ribavirin directly affected the ability of virus to infect cells, macrophages were pretreated with ribavirin for 2 h before addition of MHV-3, following which cells were cultured for an additional 10 h, harvested, and assayed for viral titers. There was no significant difference on inhibition of viral replication in the presence or absence of preincubation suggesting that the inhibitory effect was not due to inhibiting entry of MHV-3 into the cell (Fig. 1).
Macrophages infected with MHV-3 for 8 h demonstrated a significant rise in PCA (941 ± 80 mU/106 cells) in comparison with basal values of 63 ± 15 mU/106 cells. With ribavirin treatment over the range 100 to 500 μg/ml, macrophages PCA decreased significantly from 941 ± 80 mU/106 cells to 34 ± 11 mU/106 cells (maximum 94% inhibition) (p < 0.001). The inhibitory effect of ribavirin showed a concentration-dependent manner (Fig. 2). To determine whether ribavirin inhibited the one-stage clotting assay, ribavirin (500 μg/ml) was added to MHV-3-infected macrophages just before the PCA assay. Ribavirin had no inhibitory effect on the ability to determine PCA (data not shown).
Effect of ribavirin on BALB/cJ macrophage PCA induced by MHV-3. One million macrophages were stimulated with MHV-3 at a MOI of 2.5 in the presence or absence of ribavirin and harvested after 8 h for measurement of PCA activity. Values represent the mean ± SD of three separate experiments done in triplicate. *represents p < 0.01 when compared with MHV-3-stimulated macrophages.
Northern blot analysis showed MHV-3-induced mRNA of fgl-2 was inhibited by ribavirin. This effect was dose dependent with inhibition seen at 100 μg/ml and complete inhibition observed at 500 μg/ml of ribavirin, as seen in Figure 3.
Effect of ribavirin on transcription of mRNA of fgl2 in MHV-3-infected BALB/cJ macrophages. Thirty micrograms of total RNA extracted from macrophages that had been stimulated with MHV-3 at a MOI of 2.5 were added to each lane and hybridized with musfiblp cDNA (E1B) as described in Materials and Methods. Lane 1, macrophages alone; Lane 2, macrophages + ribavirin (500 μg/ml); Lane 3, macrophages + MHV-3; Lane 4, macrophages + MHV-3 + ribavirin (500 μg/ml); Lane 5, macrophages + MHV-3 + ribavirin (100 μg/L); Lane 6, macrophages + MHV-3 + ribavirin (1 μg/ml). A glyceraldehyde-3-phosphate dehydrogenase (GAPDH) cDNA was used to ensure equal amounts of RNA in all lanes.
Macrophages produced significantly higher levels of TNF-α in response to MHV-3 infection (10.73 ± 2.15 ng/ml) in comparison with the basal values (0.03 ± 0.03 ng/ml). Ribavirin significantly inhibited the production of TNF-α in MHV-3-infected macrophages over the range 100 to 500 μg/ml. The inhibitory effect showed a dose-dependent pattern with a maximum 75% inhibition in TNF-α (10.73 ± 2.15 to 2.74 ± 0.93 ng/ml) (p < 0.001) (Fig. 4). By Northern blot analysis, addition of ribavirin prevented increased levels of TNF mRNA transcription seen in response to MHV. The reduced effect was first detected at a concentration of 100 μg/ml with complete inhibition at 500 μg/ml (Fig. 5 A).
Effect of ribavirin on BALB/cJ macrophage TNF-α production induced by MHV-3. One million macrophages from BALB/cJ mice were stimulated with MHV-3 at a MOI of 2.5. TNF levels in the supernatants were measured by ELISA after incubation for 8 h. Data are presented as mean ± SD for three separate experiments done in duplicate. * represents p < 0.01 compared with macrophages + MHV-3.
Effect of ribavirin on mRNA level of TNF-α and IL-1β in BALB/cJ macrophages stimulated with MHV-3 by Northern blot analysis. Thirty micrograms of total RNA extracted from macrophages that had been stimulated with MHV-3 at a MOI of 2.5 were added to each lane and hybridized with TNF-α cDNA (A) or IL-1β cDNA (B) as described in Materials and Methods. Lane 1, macrophages alone; Lane 2, macrophages + ribavirin (500 μg/ml); Lane 3, macrophages + MHV-3; Lane 4, macrophages + MHV-3 + ribavirin (500 μg/ml); Lane 5, macrophages + MHV-3 + ribavirin (100 μg/L); Lane 6, macrophages + MHV-3 + ribavirin (1 μg/ml). A glyceraldehyde-3-phosphate dehydrogenase (GAPDH) cDNA was used to ensure equal amount of RNA in all lanes.
Supernatants from macrophages were analyzed for production of IL-1. The dose-dependent inhibitory effect of ribavirin on the IL-1β production of MHV-3-stimulated macrophages is depicted in Figure 6. Ribavirin significantly inhibited IL-1β production from 155.91 ± 22.62 to 5.74 ± 0.70 pg/ml over the range 10 to 500 μg/ml (maximum 96% inhibition) (p < 0.001). This inhibitory effect was at the level of gene transcription as evidenced by Northern blot analysis (Fig. 5 B). The mRNA concentration for IL-1 was inhibited by ribavirin (100 μg/ml) and was undetectable when cells were treated with 500 μg/ml ribavirin.
Effect of ribavirin on BALB/cJ macrophage IL-1β production induced by MHV-3. One million macrophages from BALB/cJ mice were stimulated with MHV-3 at a MOI of 2.5. IL-1β levels in the supernatants were measured by ELISA after incubation for 8 h. Data are presented as mean ± SD for three separate experiments done in duplicate. * represents p < 0.01 compared with + MHV-3.
To determine whether the inhibitory effect of ribavirin on transcription of mRNA for IL-1 and TNF was specific or a general phenomenon, the ability of ribavirin to inhibit transcription of mRNA for these cytokines following stimulation with endotoxin (LPS) was determined. Ribavirin had no effect on mRNA levels of TNF-α and IL-1 in LPS-stimulated macrophages even when added at concentrations up to 500 μg/ml (Fig. 7).
Effect of ribavirin on mRNA levels of TNF-α and IL-1β in BALB/cJ macrophages stimulated with LPS by Northern blot analysis. Thirty micrograms of total RNA extracted from macrophages that had been stimulated with LPS at 10 μg/ml added to each lane and hybridized with TNF-α cDNA (A) or IL-1 cDNA (B) as described in Materials and Methods. Lane 1, macrophages alone; Lane 2, macrophages + ribavirin (500 μg/ml); Lane 3, macrophages + LPS; Lane 4, macrophages + LPS + ribavirin (500 μg/ml); Lane 5, macrophages + LPS + ribavirin (100 μg/l); Lane 6, macrophages + LPS + ribavirin (1 μg/ml). A glyceraldehyde-3-phosphate dehydrogenase (GAPDH) cDNA was used to ensure equal amounts of RNA in all.
In vitro.
To determine whether ribavirin differentially affected production of Th1/Th2 cytokines, ribavirin (1–500 μg/ml) was added to the T cell lines. Supernatants were collected and assayed for the production of IFN-γ and IL-4 as described above. Ribavirin, when added at concentrations from 1 to 500 μg/ml, had no inhibitory effect on the production of IFN-γ (84.5 ± 3.7 ng/ml) by either the Th1/Th2 line or the Th1 cell line 3E9.1, as shown in Table I. Addition of ribavirin at a concentration higher than 100 μg/ml inhibited the production of IL-4 by both the Th1/Th2 line and the Th2 line 4B6.8 (Table I). A maximum 97% inhibition (p < 0.01) was seen at concentrations of ribavirin ≥100 μg/ml and was not due to toxic effect of ribavirin on Th2 cells as shown by trypan blue exclusion.
Effect of ribavirin on cytokine production by T cell linesa
Ribavirin (μg/ml) .
IFN-γ (ng/ml) .
IL-4 (U/ml) .
Th1/Th2 .
3E9.1 .
4B6.8 .
0 85 ± 5 114 ± 12 5.6 ± 2.8 16 ± 1 0.3 ± 0.1 18.3 ± 1.6
1 85 ± 5 114 ± 12 4.1 ± 1.1 16 ± 1 0.3 ± 0.16 12.2 ± 1.1
10 76 ± 7 110 ± 10 5.2 ± 1.2 11 ± 4 0.2 ± 0.10 8.1 ± 2.0*
100 81 ± 5 110 ± 10 5.1 ± 1.1 1 ± 1* 0.1 ± 0.10 0.4 ± 1.0*
500 78 ± 8 114 ± 12 4.8 ± 1.2 0.5 ± 0.1* 0.2 ± 0.10 0.1 ± 0.1*
The Th1/Th2 cell line was derived from C3H/HeJ mice. 3E9.1 is a Th1 cell line derived from A/J mice as previously described (8). 4B6.8 is a Th2 cell line derived from BALB/cJ mice as previously described (8). Th1/Th2, Th1 (3E9.1), and Th2 (4B6.8) cells (2 × 104) were cultured with various concentrations of ribavirin for 24 h. IFN-γ and IL-4 were measured in the supernatants as described in Materials and Methods. Results represent the mean ± SD of three experiments done in triplicate. (* indicates p < 0.001 compared with no ribavirin treatment.
In vivo.
To determine the effect of ribavirin on levels of IL-4 and IFN-γ in vivo, mice fully susceptible (BALB/cJ), semisusceptible (C3H/HeJ), and resistant (A/J) to MHV-3 were treated daily with either ribavirin at a dose of 75 mg/kg/day i.p. or saline beginning 2 h before infection with 100 PFU of MHV-3. Groups of mice (n = 4) were sacrificed daily, and sera were collected and assayed for IL-4 and IFN-γ. Similar to previous results (21), sera from MHV-3-infected A/J mice had an increase in levels of IFN-γ, but no increase in levels of IL-4 consistent with a Th1 response whereas sera from MHV-3-infected BALB/cJ mice had an increase in IL-4, but no increase in IFN-γ consistent with a Th2 response. Sera from MHV-3-infected semisusceptible C3H mice had a cytokine profile similar to that of susceptible BALB/cJ mice. Treatment of mice with ribavirin resulted in a marked inhibition in levels of IL-4 in semisusceptible and susceptible mice, whereas it had little effect on production of IFN-γ in resistant mice similar to the observed effects in vitro in the Th1, Th2, and Th1/Th2 lines (Table II).
Effect of ribavirin on cytokine release following MHV-3 infection in vivoa
Mouse Strain .
Treatment Protocol .
Saline .
Ribavirin .
IL-4 (pg/ml) .
BALB/cJ 28.4 ± 4.1 2.8 ± 1.1 6.2 ± 1.1* 5.5 ± 2.0
A/J 2.3 ± 1.1 171 ± 11 2.1 ± 1.2 156 ± 8
C3H/HeJ 22.4 ± 8.2 15.6 ± 2.4 3.7 ± 2.1* 19 ± 3
Concentrations of IL-4 and IFN-γ were measured on day 3 post-MHV-3 infection in sera and are mean ± SD from four mice per group. IL-4 and IFN-γ measured as described in Materials and Methods. All mice infected with 100 PFU of MHV-3. * P <0.01 compared with saline-treated mice.
Viral hepatitis is a major health problem accounting for significant morbidity and mortality. An incomplete understanding of the pathogenesis of viral hepatitis has limited the development of successful medical approaches to its treatment (14). Recent studies have suggested that the antiviral agent ribavirin alone or in combination with IFN may be of benefit in the treatment of patients with hepatitis B (15) and C (1, 16, 17, 18). However, the mechanism for the beneficial effect of ribavirin remains unknown given that ribavirin appears not to eradicate viral replication. Our laboratory has been interested in determining the mechanism for susceptibility to viral hepatitis and has extensively examined the immunopathogenesis of viral hepatitis using an experimental animal model of acute and chronic liver disease induced by mouse hepatitis virus strain 3 (MHV-3) (8, 19, 20, 21, 22). Studies to date have suggested that resistance to MHV-3 is associated with a predominant Th1 response, the production of IFN, neutralizing Abs, and cytotoxic T cells (8, 22). In contrast, in susceptible mice, viral infection of macrophages leads to a marked inflammatory response including sustained production of TNF, IL-1, a unique procoagulant fgl2 prothrombinase encoded by the gene fgl2 located on mouse chromosome 5, and is associated with a Th2 cellular immune response and production of nonneutralizing Abs (23). Previous studies in our laboratory have demonstrated that susceptibility to MHV-3 correlates with macrophage activation and not viral replication (20). The importance of the fgl2 prothrombinase, which can directly cleave prothrombin to thrombin, has been demonstrated by the fact that treatment of mice with a high titered neutralizing Ab to fgl2 prothrombinase fully protected these mice from the lethality of MHV-3 (6). In contrast, inactivated virus (MHV-3) did not induce the production of these inflammatory mediators and did not cause disease (24). Previous studies have shown that ribavirin prolonged the course of MHV-3 infection and increased the survival of animals (1, 4). The present studies were undertaken to determine the mechanism of the beneficial effects of ribavirin in the experimental animal model of MHV-3.
Our data confirm that although ribavirin has minimal (<1 log) inhibitory effects on replication of MHV-3 in vitro, even at very high concentrations complete eradication of the virus was not seen, as has been previously reported (1, 25, 26). However, ribavirin at concentrations that are achieved in vivo (4) almost totally inhibited the production of proinflammatory mediators TNF, IL-1, and PCA in BALB/cJ macrophages in vitro. Ribavirin was unable to cause similar inhibition of LPS-induced inflammatory cytokines. This may be due to the fact that induction of TNF, IL-1, and PCA is due to LPS induction of IFN γ which is not inhibited by ribavirin (27, 28).
In several other animal models of liver injury, including those due to virus infection, endotoxin, CCl4, galactosamine, and acetaminophen, the hepatic injury is associated with fibrin deposition, sinusoidal thrombosis, and accumulation of the inflammatory cells (29, 30, 31, 32, 33, 34, 35, 36, 37). In the hepatocellular necrosis associated with these pathologic processes, resident macrophages within the liver (Kupffer cells) exhibit morphologic features of activation and release a number of inflammatory mediators, including TNF, IL-1, proteolytic enzymes, and eicosanoids, as well as superoxide anions and nitric oxide (36). Furthermore, Chisari (38), using a hepatitis B surface Ag transgenic mouse model, has shown that although CD8+ CTL initiates hepatocyte injury, macrophages and their inflammatory mediators, in particular IL-1 and TNF, are responsible for massive hepatic necrosis. Inactivation of these macrophages prevents hepatic necrosis.
Macrophages generate a wide range of mediator molecules which may contribute either directly or indirectly to the development of fulminant hepatitis by inducing PCA (37). TNF and IL-1 production by macrophages can stimulate endothelial cell production of immune coagulants and increase neutrophil-endothelial interaction, thereby potentially promoting microvascular thrombosis (39, 40).
The immune system plays an essential role in the outcome of viral infection. One mechanism of the immune response in vivo involves CD4+ Th cells, which, through the production of cytokines, control the development of immune effector mechanisms such as Ab production, generation of cytotoxic T cells, and macrophage activation. Following Ag exposure, Ag-specific Th cells differentiate along two pathways (41). Th1 cells, through the production of IL-2, IFN-γ, and lymphotoxin, mediate cellular immunity, which is essential for clearance of viral and other pathogens (42, 43, 44, 45, 46, 47, 48, 49, 50, 51). Th2 cells, conversely, produce IL-4, IL-10, and IL-13 and are most effective in providing help for B cell differentiation to plasma cells. IFN-γ production by Th1 cells inhibits the proliferation of Th2 cells (43), whereas treatment with neutralizing Abs to IFN-γ promotes the development of a Th2 response (44, 45).
Our data also showed that ribavirin diminished IL-4 production both by the Th1/Th2 line and the MHV-3-specific Th2 cell line 4B6.8, while no effect was seen on IFN-γ production by Th1 cells, thus preventing the shift to a Th2 response.
In vivo, treatment of MHV-3-infected semisusceptible C3H/HeJ and susceptible BALB/cJ mice with ribavirin resulted in inhibition of IL-4 production, but similar doses of ribavirin did not inhibit production of IFN-γ production in resistant A/J mice. This may explain the benefit of ribavirin on amelioration of MHV-3 infection and the increased survival in vivo previously reported (4).
A Th1 response has been associated with host resistance, and a Th2 response has been associated with susceptibility in murine models of leishmaniasis, candidiasis, listeriosis, and MHV-3 (46, 47). Recently, in acute and chronic hepatitis B virus infection in humans, a Th1 response has also been associated with resistance and a Th2 response with susceptibility (38, 48). Furthermore, immunomodulatory treatments that have converted a Th1 to a Th2 response have resulted in the loss of resistance (38), and, conversely, induction of a Th1 response has led to resolution of the infection (48). In hepatitis C, studies on cytokine profiles have been reported only in patients with chronic disease in contrast to the model of acute fulminant hepatitis induced by MHV-3. The reports in patients with HCV are somewhat conflicting but suggest a poor or absent Th1 response in these patients (49, 50, 51). Recent studies have shown that ribavirin has beneficial effects on serum aminotransferase concentrations and necroinflammatory activity of liver biopsies in patients with chronic HCV infection. However, ribavirin has no effect on serum HCV RNA titers, and biochemical relapse is universal after cessation of therapy (15, 16, 17). The discordance between the virologic and biochemical responses in patients receiving ribavirin suggests that the beneficial effect of ribavirin may not be mediated by inhibition of viral replication. Another possibility is that ribavirin modulates the immune response to HCV.
In conclusion, our study suggests that ribavirin is a very potent inhibitor of viral-induced proinflammatory mediators. The beneficial effects of ribavirin in both experimental animal models and clinical treatment may be related to its ability to markedly reduce macrophage activation and diminish Th2 cytokine production while preserving Th1 cytokine production. These data provide a rational potential clinical utility of ribavirin either alone or in combination with IFN-α in patients with hepatitis or as a substitute in IFN-α-nonresponsive patients.
We thank Mrs. Charmaine Beal for preparation of the manuscript.
This work was supported by a group grant (PG11810) from the Medical Research Council of Canada.
Abbreviations used in this paper: MHV-3, mouse hepatitis virus strain 3; PCA, procoagulant activity; musfiblp, mouse fibrinogen-like protein; HCV, hepatitis C virus; PFU, plaque-forming units; MOI, multiplicity of infection.
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. Reciprocal expression of interferon gamma or interleukin 4 during the resolution or progression of murine leishmaniasis: evidence for expansion of distinct helper T cell subsets.
Scott, P..
. IFN-γ modulates the early development of Th1 and Th2 responses in a murine model of cutaneous leishmaniasis.
Belosevic, M., D. S. Finbloom, P. H. Van der Meide, M. V. Slayter, C. Nacy.
. Administration of monoclonal anti-IFN-gamma antibodies in vivo abrogates natural resistance of C3H/HeN mice to infection with Leishmania major.
Swain, S. L., A. D. Weinberg, M. English, G. Huston.
. IL-4 directs the development of Th2-like helper effectors.
Chatelain. R., K., K. Varkila, R. L. Coffman.
. IL-4 induces a Th2 response in Leishmania major-infected mice.
Romani, L., A. Mencacci, U. Grohmann, P. Mosci, P. Puccetti, F. Bisstoni.
. Neutralizing antibody to interleukin 4 induces systemic protection and T helper type 1-associated immunity in murine candidiasis.
Modlin, R. L., T. B. Nutman.
. Type 2 cytokines and negative immune regulation in human infections.
Schlaak, J. F., T. Baecker, H. F. Lohr, K.-H. Meyer zum Buschenfelde.
. Chronic hepatitis C: IL-12 enhances HCV-antigen induced cytokine production of PBMC in a subgroup of patients.
Lechmann, M., B. Langhans, S. Bar, R. Woitas, L. Leifeld, H. G. Ihlenfeldt, G. Jung, W. Effenberger, T. Sauerbruch, U. Spengler.
. Deficient secretion of TH1 cytokines in patients with chronic hepatitis C.
Woitas, R. P., M. Lechmann, B. Langhans, S. Bar, H. G. Ihlenfeldt, G. Jung, T. Sauerbruch, U. Spengler.
. Cytokine response to hepatitis C virus core peptides in patients with chronic hepatitis C infection and healthy anti-HCV seropositives.