[The prevalence of hepatitis C virus (HCV) subtypes in Chinese HIV-1/HCV co-infected individuals].

OBJECTIVE: To better understand the prevalence and geographic distribution of genotypes/subtypes on HCV and the relationship between HCV genotypes/subtypes and HIV infection disease progression in the HIV-1/HCV co-infected individuals living in high HIV-1 prevalent areas in China. METHODS: 186 plasma samples were collected from HIV-1 seropositive individuals infected through paid blood donors (PBD), injecting drug users(IDUs) or sexual contact, living in most severely affected provinces, Henan, Yunnan, Xinjiang, Jilin and Liaoning provinces. Samples with HCV viral load >1000 cop/ml were amplified by RT-nested PCR, sequenced and phylogenetically analyzed for genotyping/subtyping of HCV. HIV-1, HCV viral loads and CD4+ T lymphocytes were measured for all subjects. RESULTS: (1) HCV were identified as 1a (1.7%), 1b (39.9%), 2a (17.9%), 3a (10.4%), 3b (15.6%), 6a (1.2%), 6n (6.4%), and a newly unclassified subtype (7.5%). HCV 2a and 1b subtypes predominated in PBD in Henan, 3a and 3b in IDUs in Xinjiang and Yunnan, and 6 genotype/subtypes in IDU in Yunnan. (2) There were no significant differences in CD4+ T cell counts among the different HCV subtypes. (3) The viral load of HCV RNA in 1b subtype was higher than that of non-1b subtype, however, no significant differences in HIV-1 viral loads and CD4+ T cell counts were found between 1b and non-lb subtype. Both HIV and HCV viral loads were lower in 2a than non-2a subtype. CONCLUSION: The prevalence of HCV genotype/subtype in HIV-1/HCV co-infected individuals was associated with geographic areas and transmission routes. HCV subtypes had no direct correlation with HIV infection disease progression.

[Frequency of the CD4+CD25nt/hiCD127lo regulatory T lymphocytes from the peripheral blood in Chinese healthy individuals].

AIM: To determine the frequency of the CD4(+)CD25(nt/hi)CD127(lo) regulatory T lymphocytes from the peripheral blood in the Chinese healthy individuals and provide some useful evidence for clinical research of correlative diseases. METHODS: From the CD4(+)CD25(nt/hi)CD127(lo) regulatory T lymphocytes of peripheral blood in 312 Chinese healthy male and female individuals aged from 8 to 60(five age groups were collected) The expression of transcription factor Foxp3 was detected by triplex immuno fluorescence and the frequency of CD4(+)CD25(nt/hi)CD127(lo) regulatory T lymphocytes was determined by flow cytometry. RESULTS: The frequency of CD4(+)CD25(nt/hi)CD127(lo) regulatory T lymphocytes in Chinese healthy individuals was (6.55+/-0.11)%, and the frequency differed among age groups(P=0.015) and sex groups(P<0.05). CD4(+)CD25(nt/hi)CD127(lo) regulatory T lymphocytes specifically express transcription factor Foxp3. CONCLUSION: The frequency of the CD4(+)CD25(nt/hi)CD127(lo) regulatory T lymphocytes from the peripheral blood in the Chinese healthy individuals has been preliminarily determined which lays the foundation for further clinical research of regulatory T lymphocytes. As a specific cell surface marker, CD25(nt/hi)CD127(lo) can helpful obtain pure CD4(+)CD25(+) regulatory T lymphocytes and suppress the interference of other cells during cell separation.

Genetic diversity and drug resistance of human immunodeficiency virus type 1 (HIV-1) strains circulating in Shanghai.

The HIV-1 epidemic in Shanghai is rapidly increasing. To better understand the HIV-1 genetic diversity and the mutations associated with resistance to protease inhibitors (PIs) and reverse transcriptase inhibitors (RTIs), 95 antiretroviral (ARV)-treated and treatment-naive HIV-1-seropositive individuals living in Shanghai were investigated. The HIV-1 pol gene in 70 of the 95 plasma samples was successfully amplified and analyzed. The result showed that CRF01_AE predominated in Shanghai with 42.9%, followed by subtype B (10%), B' (12.9%), CRF07_BC (11.4%), CRF08_BC (10%), CRF02_AG (4.3%), G (2.9%), and K (1.4%). In addition, three new intersubtype and/or inter-CRF recombinants were detected including B'/CRF01_AE (1.4%), U/G (1.4%), and U/CRF01_AE (1.4%). The mutations conferring primary and secondary resistance to PIs were detected in 3 of 70 (4.3%) patients and the mutations conferring resistance to RTIs were identified in 12 of 70 (17.2%) patients, among whom 11 of 15 (73.3%) and 1 of 55 (1.8%) were ARV-treated and treatment-naive individuals, respectively (p < 0.01). This study reveals the emergence of genetic diversity of HIV-1 currently circulating in Shanghai. HIV-1 infection by heterosexual contact is still a major route for introduction of HIV-1 variants into this city in recent years. It is believed that this information may help to guide recommendations for diagnostic assays, vaccine design, and antiretroviral regimen strategies in China.

Efficient delivery of human clotting factor IX after injection of lentiviral vectors in utero.

AIM: To explore gene transfer feasibility for human clotting factor IX (hFIX) mediated by recombinant lentivirus in utero. METHODS: ICR mice fetus at 17-19 d gestation were received lentiviral vectors carrying hFIX cDNA under the control of liver specific promoter by intrahepatic injection. The expression and distribution of hFIX cDNA and possible immune responses against the hFIX were assessed by ELISA, PCR, RT-PCR, and immunohistochemistry, respectively. RESULTS: The serum hFIX protein were detected at different time points in all newborn mice, the highest level of hFIX was 50 microg/L and lasted for more than 30 d. Anti-hFIX antibody was not detected. hFIX cDNA was detected in liver, spleen, and heart. The expression of hFIX cDNA was only detected in liver. Besides, no germ line transmission was found at DNA and RNA levels, and no side effect associated with gene transfer was detected. CONCLUSION: The efficient delivery of hFIX can be achieved by prenatal gene transfer. It thus shows the feasibility of gene therapy for hemophilia in utero.