A novel SELEX method for screening of HIV-1 P24 antigen / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To screen aptamers that can bind P24 antigen tightly and specificly, and verify its specificity and affinity.</p><p><b>METHODS</b>Polycarbonate PCR plate was coated with P24 antigen and SELEX technology was used to screen aptamer on the PCR plate. The primary and secondary structure of these aptamers was analyzed by software. Through HRP labeled streptavidin and biotin labeled aptamers, the affinity and specificity of obtained aptamers were verified by ELISA.</p><p><b>RESULTS</b>The polycarbonate PCR plate could be coated with P24 antigen. Electrophoretic analysis showed the aptamers had been enriched. Sequence aligment analysis showed that these aptamers have consensus sequence and their apatial structure was multiple; ELISA verified that aptamers had high affinity with P24 antigen.</p><p><b>CONCLUSION</b>A simple method was established for screening aptamers that can bind HIV-1 P24 antigen specificly and tightly.</p>

Reduction of the HIV Seroconversion Window Period and False Positive Rate by Using ADVIA Centaur HIV Antigen/Antibody Combo Assay

BACKGROUND: Early diagnosis of HIV infection reduces morbidity and mortality. Fourth-generation HIV detection assays are more sensitive because they can detect p24 antigen as well as anti-HIV antibodies. In this study, we evaluated the performance of a new fourth-generation ADVIA Centaur HIV antigen/antibody combo (CHIV) assay (Siemens Healthcare Diagnostics Inc., USA) for early detection of HIV infection and reduction of false positive rate. METHODS: Four seroconversion panels were included. The third-generation ADVIA Centaur HIV 1/O/2 enhanced (EHIV) assay (Siemens Healthcare Diagnostics Inc., USA) and fourth-generation CHIV assay were used to test each panel for HIV infection. The presence of antigen was confirmed using HIV p24 antigen assay. To evaluate false-positivity and specificity, 54 HIV false-positive and HIV-negative serum samples from 100 hospitalized patients and 600 healthy subjects were included. RESULTS: Compared to the EHIV assay, the CHIV assay had a shorter window for three of the seroconversion panels: a difference of 10 days and two bleeds in one panel, and 4 days and one bleed in the other two panels. Only 34 of the 54 (63%) samples known to yield false-positive results by EHIV assay had repeatedly yielded reactive results in the CHIV assay. One of the 600 healthy subjects had a false-positive result with the CHIV assay; thus, the specificity was 99.85% (699/700). CHIV accurately determined the reactive results for the HIV-confirmed serum samples from known HIV patients and Korea Food & Drug Administration (KFDA) panels. CONCLUSIONS: The new fourth-generation ADVIA Centaur HIV assay is a sensitive and specific assay that shortens the serological window period and allows early diagnosis of HIV infection.

Anti-HIV-1 activity and structure-activity relationship of pyranocoumarin analogs / 药学学报

The purpose of this study is to find out anti-HIV-1 reverse transcriptase (RT)/protease (PR) activity and inhibition of virus replication in cell cultures of novel coumarin analogs and determine their structure-activity relationship. Coumarin derivatives have been demonstrated to inhibit the activity of HIV-1 RT/PR in cell free system. It also shows inhibition effects to HIV-1 replication in cell culture. Based on the Chinese traditional pharmacological characteristics and protein three dimension computer aided design, analogs of tetracyclic dipyranocoumarin were synthesized from natural leading compounds. We studied the relationship of antiviral effects and chemical structures via HIV-1 PR/RT enzyme models and cell culture model system. Seven compounds were designed and tested. Several compounds showed anti-HIV-1 activity in varying degrees, especially V0201 showed much higher anti-HIV-1 activity with 3.56 and 0.78 micromol x L(-1) of IC50 against HIV-1 PR/RT and 0.036 micromol x L(-1) against HIV-1 replication in PBMC cultures. V0201 with a novel structure may be a new leading compound. These new compounds are valuable for development of new anti-HIV drugs in the future.

Study on vector-specific and foreign gene-specific immune responses induced by rAd5 and rAAV2/1 vaccines / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To compare the foreign gene-specific and vector-specific immune responses in BALB/c mice immunized with rAd5 or rAAV2/1 expressing the same gene.</p><p><b>METHODS</b>BALB/c mice were immunized with rAd5-gag or rAAV2/1-gag once, HIV-1 Gag-specific and vector-specific cellular immune responses were analyzed by Elispot assay, HIV-1 P24-specific IgG and vector-specific IgG were tested by ELISA assay.</p><p><b>RESULTS</b>Mice immunized with rAd5-gag induced potent Gag-specific cellular immune responses and that were significantly higher than Ad5-specfic cellular responses, while rAAV2/1-gag elicited weak Gag-specific and AAV2/1-specific cellular responses. Both P24-specific and Ad5-specific IgG titers induced by rAd5-gag were high and in similar level. Higher level of P24-specific IgG was found in mice inoculated with rAAV2/1-gag than rAd5-gag. And the P24-specific IgG titers were higher than the vector-specific IgG titers in mice immunized with rAAV2/1.</p><p><b>CONCLUSION</b>rAd5 could elicit strong foreign gene-specific cellular and humoral immune responses, weak vector-specific cellular responses and strong vector-specific antibodies, rAAV2/1 could induce potent foreign gene-specific antibodies that were much higher than vector-specific IgG, while both foreign gene-specific and vector-specific cellular responses were very low.</p>

Immunological efficiency induced by HIV-1 p24 DNA combined with P24 protein / 生物工程学报

New strategies to improve vaccine efficacy against human immunodeficiency virus type 1 (HIV-1) are still required. DNA vaccines, exhibiting potential advantages over conventional vaccines for their simplicity and versatility, can induce specific humoral and cellular immune responses. We developed a recombinant pVAX1 DNA vector carrying p24 gene of HIV-1. The results showed that pVAX1 mediated gene possessed the ability of effective expression in both transfected 293T cells and BALB/c mice. And pVAX1-p24 DNA prime and boost immunization can induce significant P24-specific humoral immune responses and cellular immune responses in BALB/c mice. Furthermore, immunization with pVAX1-p24 DNA prime and protein boost induced 7.3 to 8.0-fold greater p24-specific humoral responses than pVAX1-p24 DNA prime and boost, while the cellular immune responses induced by combined immunization was lower. The results suggested that pVAX1-p24 DNA and P24 protein vaccine is a promising HIV-1 vaccine, and the selections of the immunization strategies are important for the immunization results.

Inhibition of the replication of HIV-1 by norcantharidin in vitro / 药学学报

For obtaining new structural compounds with unique resistance profiles or novel mechanisms of action on HIV-1 from natural products, anti-HIV-1 drug screening models were used in vitro. Norcantharidin (NCTD), a derivative from cantharidin, was found to have inhibitory activities on HIV-1(IIIB) p24 antigen in lymphocyte lines MT-4, CEM and H9. It inhibited HIV-1 strain 018a (sensitive to zidovudine) from replicating with EC50 (50% effective concentration) of 14.9 micromol L(-1) and also inhibited HIV-1 strain 018c (resistant to zidovudine) from replicating with EC50 of 20.2 micromol L(-1) in primary lymphocytes peripheral blood mononuclear cells (PBMC). Norcantharidin showed synergistic activity with zidovudine on HIV-1(IIIB) in MT-4 cells, the combination index was less than 0.3. But, it was not active on HIV-1 integrase, reverse transcriptase or protease in vitro. As the structure of norcantharidin is unique and different from that of all clinic drugs approved, it would be possible to obtain new and effective compounds against HIV-1 with low toxicities after modification of norcantharidin.

Assessment of a self-deferral form for screening blood donors, Chiang Mai University Hospital, Thailand.

A self-deferral form has been used to screen Chiang Mai University Hospital blood donors and was improved in 2005. It has never been evaluated. The study aimed to assess the self-deferral form procedures in detecting infected donors. Sera from 5,083 donors, who passed the self-deferral screening form, were tested with the routine immuno-assays (serology) for HIV 1 and 2 antibodies, P24 antigen, HCV antibodies, HBV surface antigen, and syphilis. Antibody negative sera were also tested individually with the the Procleix Ultrio Assay for HIV-1 DNA, HCV RNA, and HBV DNA. The donors who had discrepant results between serology and NAT were evaluated with additional tests, including a more sensitive Alternative Nucleic Acid Test, AntiBcore IgM, AntiBcore IgG, HBsAg and Anti HBs. Among 5,083 donors, 331 (6.5%) had at least one positive marker. In multiple logistic regression analysis, the statistically significant factors (adjusted odds ratio and 95% CI) for infection were age 30 years or below [1.45 (1.03, 2.03)], male gender [2.73 (1.64, 4.56)], primary school or lower education [1.56 (1.09, 2.23)], first-time donation [1.82 (1.25, 2.67)], and frequent donation [0.80 (0.70, 0.92)]. The safest donors were females, older than 30 years, with an education more than primary school, and frequent donation. Because of missing responses to some sensitive questions, there remains a need for further improvement of the self-deferral form.

Evaluation of HIV-1 P24 antigenemia in HIV seronegative antenatal attendees in two Nigerian hospitals

Pregnant women who a tended antenatal clinics in Asokoro District Hospital. Abuja in Northern Nigeria and Tnivcrsik of Calabar Peaching Hospital. Calabar in Southern Nigeria were recruited tor this study. The subjects were aged 20 years to 40 years old. Samples collected were analysed for P24 antigenemia using the method of Niel based on the principle of Enzyme linked Immunosorbent Assay [RL1SA]. Samples were screened for HIV antibodies using the ELISA technique of imrmmocomb test using the method of Epstein. Serology testing is the routine screening method for HIV in virtually all Nigerian health centers. Two hundred and twenty pregnant women selected for this study were tested sero-negative for HIV-1 antibodies while 30 that had been tested zero positive for HIV-1 antibodies were used as controls for P24 antigenemia testing. The 220 that were seronegative for HIV-1 antibodies were tested for P24 antifiens. Thirty two [14.5%] of the 220 subjects were positive for P24 antigenemia. Out of these 32. twenty three [23] were in the highly reproductive and sexually active age brackets of 20 years to 30 years. Expectedly. 26 [86.7%] of the 30 controls tested positive for P24 antigenemia. Subjects from polygamous marriages had a higher number of P24 anligenemia positivity than those from monogamous marriages [Table 4]. This implies that mother to child transmission of HIV infection is high in sexually active reproductive age bracket and in polygamous marriages. The latter could be due to multiple partnership involvement. Observation is that using HIV-1 serologic technique, there is about 1 in 10 ratio chance of a mother transmission of HIV to her unborn child since the window period is not covered by this technique. Although HIV antibody tests are the most appropriate, rapid and efficient means of detecting infection but P24 antigen assay is an alternative technology for diagnosing early infection before a level of detectable antibodies is reached. P24 antigen is detectable in a predictable manner between 14 to 29 days of infection. We conclude that HIV serologic test, which is used routinely and generally in most Nigerian health centers, is not enough to elucidate HIV serostatus of individuals, especially those who appear to be sero-negative Therefore introduction of P24 antigen assay, to cover greater part of the window period of the infection will not only protect the mother but also the unborn child

Anti-HIV activities of Achyranthes bidentata polysaccharide sulfate in vitro and in vivo / 药学学报

Achyranthes bidentata polysaccharide sulfate (ABPS) was a sulfated derivate derived from Achyranthes bidentata polysaccharide (ABP) which was isolated and identified from Chinese herb Achyranthes bidentata. The anti human immunodeficiency virus type 1 (HIV-1) activities were studied in vitro and in vivo. ABPS was found to inhibit HIV-1 reverse transcriptase and integrase with the 50% inhibiting concentration (IC60) of (2.948 +/- 0.556) micromol x L(-1) and (0.155 +/- 0.030) micromol x L(-1), respectively, but the parent compound ABP was not effective. ABPS inhibited HIV-1 P24 antigen with IC50 of (0.082 +/- 0.044) micromol x L(-1) and selective index (SI) of > (358 +/- 148) in MT-4 cell cultures acutely infected with HIV-1 IIIB virus, and with IC50 of (11.80 +/- 5.90) micromol x L(-1) and SI of > (24.2 +/- 12.1) in PBMC cell cultures acutely infected with clinical isolated zidovudine resistant HIV-1 virus, but there was no activity even at its concentration of 500 micromol x L(-1) in latent infection of H9/HIV-1 IIIB cell cultures. 5% sera taken from rats after intraperitoneal injection from rats with ABPS 125 mg x kg(-1) once or mice with 3 mg x kg(-1) qd for 20 days effectively inhibited HIV-1 P24 in MT-4 cell cultures, but those had no inhibitory effect when given orally. The results suggested that ABPS is a promising HIV-1 inhibitor, active on HIV-1 reverse transcriptase, integrase in vitro and HIV-1 P24 antigens in cell cultures, it was well absorbed by intraperitoneal injection but poor in oral bioavailability. It warrants further study.

Overexpression, purification of recombinant HIV-1 gp41 protein and detection of HIV antibody in urine / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To establish a specific and sensitive Enzyme-linked immunosorbent Assay (ELISA) kit for detection of HIV-1 antibody in urine using Escherichia coli expression products as coating antigen.</p><p><b>METHODS</b>The truncated HIV-1 gp41 gene fragment of major antigenic epitopes was inserted into the plasmid pET22b to obtain expression plasmid pET22b-mgp41. The recombinant antigen was expressed in BL21 (DE3) strains of Escherichia coli and was purified by immobilized metal chelation and gel filtration chromatography. Using this antigen as coating antigen, a HIV-1 urine antibody ELISA kit was developed. In order to examine the clinical utility of the kit, 5437 urine samples were assayed, which consisted of 641 urine samples from HIV infected patients and 4796 samples from normal subjects. Results The purity of purified antigen is up to 95%. Anti-HIV antibodies were detected in all the urine samples from HIV infected patients, and the diagnostic sensitivity for HIV-1 infection was 100%. In healthy control group, 71 cases showed false positive, the specificity was 98.52%.</p><p><b>CONCLUSION</b>The HIV-1 urine antibody kit can be used in screening and diagnosing for HIV-1 infection.</p>

Comparative evaluation of Hebei HIV-1 p24 kit for the detection of human immunodeficiency virus / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To probe into the feasibility of screening anti-HIV compounds by using HIV-1 p24 detection kit made by Hebei Medical University.</p><p><b>METHODS</b>The sensitivity, reproducibility and efficacy of the Hebei p24 kit were evaluated compared with the commercially available Vironostika HIV-1 Antigen Microelisa System (Biomerieux).</p><p><b>RESULTS</b>Hebei p24 kit had high sensitivity and good reproducibility. In vitro screening demonstrated that there was no statistically significant difference (P greater than 0.05) between these two kits in assessing anti-HIV compounds.</p><p><b>CONCLUSION</b>Hebei p24 kit could be used as an easily affordable alternative method for detection of HIV-1 in screening anti-HIV compounds.</p>

[Evaluation the immunogenesity structure of HIV recombinant protein gp41-p24]

HIV recombinant proteins can be used as immunogen and inducer of immune system. The gag and env proteins are among the most important ones that are located on internal and surface sections, respectively. In this study, we considered the immunogenisity structure of HIV recombinant protein gp41-p24, which has not yet been studied in detail before. In the present study, the HIV recombinant protein gp41-p24 was prepared by cloning methods and kept as unfolded. Using the dilution procedure, unfolded protein was changed to refolded state. The molecular weight and concentration of protein [refolded and unfolded] was measured by electrophoresis and spectrophotometer methods. The measurement of refolded protein can be estimated by native gel and circular dichroism method [CD] based on secondary structure of the protein. Immunological activity and immunogenic structure of these two proteins, based on protein type and optical density was recorded by ELISA and western blot methods. Our results showed that molecular weight of each protein was 32 KD and also they were pure. The refolded protein was observed by native gel method. In the above protein compared with the unfolded one, increased content of helix and beta strand structures and decreased random form was shown. Immune reaction with the antibodies in the serum of HIV positive control patients was observed in the standard and refolded proteins. There was no significant difference based on the protein type. Our research indicated that the HIV recombinant protein gp41-p24, after refolding, has immunogenic activity and we suggest its application as an immunogen in immunization and stimulation of immune system

Immune response analysis of immunized Balb/C mice with HIV-1 fusion p24 - gp41 genes as DNA vaccine candidate

The global HIV epidemic continues to expand and exceeding previous predictions. An effective vaccine represents the best hope to curtail the HIV epidemic. DNA vaccines induce humoral and cellular responses and mimic live vaccines without their pathogenic potential. The importance of CD8[+]CTL responses in controlling HIV and SIV viremia has led to production of a series of vaccines candidates that effectively induce these responses. It is now widely believed that an HIV vaccine strategy must stimulate both a strong humoral [antibody] as well as cell-mediated [CTL] immune response. The p24 and gp41 play many important roles in host-virus interaction and pathogenesis. These proteins are considered as attractive vaccine candidate in which their immunogenecity and immunomodulatory effects have been confirmed. In this study, a construct, pcDNA3.1Hygro- [p24-gp41], was evaluated as a DNA vaccine candidate in Balb/C mice for generation of effective cellular immune responses. For immunizing, we used dendrosome, a novel family of vehicles for transfection and therapy. IFN-gamma cytokine production and total antibody were detected by ELISA. Lymphoprolifration assay was performed by MTT test. ELISA and MTT assays confirmed that the cited p24-gp41 fusion gene is able to enhance immune responses in mice. The construct that was used in this research can be a good candidate for DNA vaccine against HIV-1, if the future complementary tests demonstrate the same trends of immunogenic responses shown in this study

Establishment and evaluation of the diagnostic kit for anti-HIV1/2 antibody and P24 antigen / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To establish and evaluate an Enzyme Immunoassay diagnostic kit combined with anti-HIV1/2 antibody and P24 antigen for shortening the examination window period of HIV infection in HIV laboratory diagnosis.</p><p><b>METHODS</b>The enzyme-linked reaction plates was coated by anti-HIV P24 monoclonal antibody and HIV 1/2 antigen. Labeling HIV1/2 antigen and anti-HIV P24 polyclonal antibody with horseradish peroxidase, setup an integrated ELISA kit for detecting anti-HIV-1/2 antibody and HIV P24 antigen, and evaluate the specificity and sensitivity of this kit.</p><p><b>RESULTS</b>The sensitivity of testing P24 antigen was up to 0.2 ng/ml. 78 serum samples of patients with AIDS, 85 serum samples of healthy people were compared with Abbott EIA kit, the coincidence was 100%. 12 051 sera from normal persons and patients were examined, the sensitivity of 100 %and specificity of 99.62 %, respectively.</p><p><b>CONCLUSION</b>The anti-HIV1/2 antibody and HIV P24 antigen can be measured at the same time using this EIA kit, while the examination window period of HIV infection is shortened. Thus, the method is suitable for laboratory diagnosis and epidemiological investigation.</p>

Evaluation of Abbott Fourth Generation HIV Antigen and Antibody Assays / 대한진단검사의학회지

BACKGROUND: In order to reduce the diagnostic window period between the time of human immunodeficiency virus (HIV) infection and serological diagnosis, new fourth generation screening assays which detect HIV p24 antigen and specific antibody simultaneously have been developed. In this study, we evaluated the performance of a new fourth generation assay. METHODS: We compared a new fourth generation assay, Architect HIV Ag/Ab combo, with another fourth generation assay AxSYM HIV Ag/Ab combo and a third generation assay, AxSYM HIV 1/2 gO for their performance. The assays were evaluated using 3 HIV seroconversion panels, 305 sera of healthy subjects and 100 sera of patients with HBsAg or anti-HCV antibodies. Within-run and total coefficient variations of the three screening assays were analyzed for the evaluation of precision. RESULTS: Architect HIV Ag/Ab combo shortened the window period by 8.7+/-2.1 days relative to AxSYM HIV 1/2 gO and 2.0+/-2.0 days relative to AxSYM HIV Ag/Ab combo in seroconversion panels. Architect HIV Ag/Ab combo presented the best performance in precision among the three reagents; total CV for positive control was 3.6%, 9.6% and 4.6% for Architect HIV Ag/Ab combo, AxSYM HIV Ag/Ab combo and AxSYM HIV 1/2 gO, respectively. Specificities of three assays were not different in this study. CONCLUSIONS: HIV Ag/Ab combined assays reduced the diagnostic window as compared to the third generation screening assays, enabling an earlier diagnosis of HIV infection. A new fourth generation assay, Architect HIV Ag/Ab combo presents a better performance than AxSYM HIV Ag/Ab combo, showing improved seroconversion sensitivity and precision.

Expression in Pichia pastoris, fermentation and purification of HIV-1 CN54 Gag antigen / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To express the Gag protein of HIV-1 strain CN54 in Pichia pastoris (P.pastoris), optimize fermentation parameters and purify Gag antigen.</p><p><b>METHODS</b>The Gag gene was subcloned into downstream of aox1 promoter of Pichia expression vector pPS1.0, an integrative vector which possesses an identical 5' untranslated region as the natural aox1 gene and employs both in vitro construction and in vivo selection for multi-copy integrants. The recombinant vector was introduced into P.pastoris strain GS115 by electroporation and selected with G418 for Gag gene integration. Super G418 resistant clones were selected and screened for Gag expression. The engineered P.pastoris was cultured to high cell density (>300 A600 Units/ml) in a 5L fermentor. Through methanol induction, the expression level of Gag reached 120 mg/L. Intracellularly expressed Gag was released by high-pressure homogenization and purified through Sepharose FF and DEAE Sepharose FF column chromatography, the purity of Gag reached up to 90%.</p><p><b>RESULTS</b>Western-blotting suggested that purified Gag expressed in P.pastoris could react specifically with serum from HIV infected individual.</p><p><b>CONCLUSION</b>Gag antigen expressed in P.pastoris has provided a good basis for the development of a new generation of HIV vaccine candidates against some Chinese prevalent strains.</p>

Immunohistochemical evidence for HIV-1 infection in the liver of HIV-infected patients / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To detect p24 antigen of human immunodeficiency virus (HIV)-1 in the liver biopsy specimens of patients with HIV infection.</p><p><b>METHODS</b>Liver biopsy samples from 14 patients with HIV/AIDS (11 man, 3 women; age range 27-52 years; infection time range 8-13 years) were examined by immunohistochemistry prospectively.</p><p><b>RESULTS</b>Intracellular expression of HIV-1 p24 antigen was detected in Kupffer cells, endothelial cells and hepatocytes. There were more HIV-positive liver cells in the patients with severer liver damage than those with milder liver damage (t=2.5189, P=0.0270).</p><p><b>CONCLUSION</b>These findings indicate that HIV-1 could replicate in the liver of HIV-infected patients and might be related to the liver cells apoptosis.</p>

Prokaryotic expression and purification of human immunodeficiency virus p24 antigen / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To express and purify the HIV p24 gene in E. coli cells, and identify p24 antigen activity.</p><p><b>METHODS</b>The full length gene fragment of HIV p24 was amplified by PCR and inserted into the pRSET vector in order to construct the pRSET-p24 recombined vector. After transforming into E. coil, the purified p24 protein was prepared by metal-ligand affinity chromatography (IMAC). The accuracy of inserted gene and activity, specificity of HIV p24 proteins were detected by two enzymes digestion technology, SDS-PAGE, Western Blot (WB) and ELISA.</p><p><b>RESULTS</b>The length of the HIV p24 gene fragment was 690 bp after digesting the recombinant plasmid T-p24 and pRSET-p24 with BamH I and Hind III. The expressed proteins had a single expected band of about 24 x 10(3) in SDS-PAGE. The specificity and activity of p24 protein were tested by WB and ELISA.</p><p><b>CONCLUSION</b>The HIV p24 sequence from HIV-1 gene plasmid hasbeen expressed in E. coil. This protein possessed good specificity and activity.</p>

A pathological study on liver tissues of patients with HIV infection / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study the pathological changes of the liver tissues of patients with HIV infection.</p><p><b>METHODS</b>14 biopsy and 12 autopsy liver tissues were examined histologically. HIV-1 related antigen of outer membrane protein gp120 and capsid protein p24 were examined with their corresponding monoclonal antibodies by immunohistochemistry.</p><p><b>RESULTS</b>In the biopsy group, cytomegalic virus (CMV) infection was found in one (1/14) case, outer membrane protein gp120 and/or capsid protein p24 antigen were detected in Kupffer cells and in some of the lymphocytes in 11 cases. All the hepatocytes were negative for outer membrane protein gp120 and capsid protein p24 antigens. In the autopsy group, there were 5 (5/12) cases of liver tissues with CMV infection and 5 cases each with mycobacterium and Toxoplasma gondii infection. Capsid protein p24 was detected in liver tissues in 3 cases.</p><p><b>CONCLUSION</b>There is HIV infection in liver tissue of patients with HIV. The rate of opportunistic infections in liver biopsy samples was lower than that in the autopsy liver tissues of patients with HIV.</p>

In vitro susceptibility of Thai seronegative donor CD8+ T lymphocytes to human immunodeficiency virus-1 (HIV-1) infection.

To determine whether CD8+ T lymphocytes from Thai donor cells are susceptible to HIV-1 infection, undepleted peripheral blood mononuclear cells (PBMC) and CD8-enriched PBMC were infected with HIV-1 Thai subtype B and CRF01_AE (E) primary isolates. Virus kinetics in HIV-1 infection of CD4+ and CD8+ T lymphocytes peaked at day 7 or 10 post infection (pi); the TCID50 used for cell infection was proportional to the level of p24 production in the cultures. We also found that the level of p24 antigen in the supernatants of infected undepleted PBMC was significantly higher than that of infected CD8-enriched PBMC. Interestingly, both single positive T lymphocytes (CD4+ and CD8+ T lymphocytes) as well as double positive CD4+/CD8+ T lymphocytes were infected with HIV-1. The double positive T lymphocytes in PBMC were found only in the presence of both CD4+ and CD8+ T lymphocytes. The majority of p24+/CD4-/CD8- T lymphocytes were HIV-1 infected CD4 down-modulated PBMC. This report provides direct evidence that single positive CD8+ T lymphocytes and double positive CD4+/ CD8+ T lymphocytes from Thai donors can be infected with HIV-1 subtypes B and E in vitro.