Serum samples from three populations of Papua New Guinea, where Plasmodium falciparum malaria and human T-lymphotropic virus type 1 (HTLV-1) are coendemic at high prevalence rates, showed statistically significant ELISA co-seropositivity and co-seronegativity. Cross-reactivity was further indicated by the presence of 10 bands ranging from 134 kDa to 18 kDa on immunoblots of electrophoresed whole lysate P. falciparum antigen against serum of HTLV-1 seropositive patients from an area where malaria is not present. Similarly, sera from patients positive for human immunodeficiency virus (HIV) from a non-malarious region produced immunoblot bands ranging from 134 kDa to 33 kDa to the P. falciparum antigen. The HTLV-1 and HIV serum samples yielded a number of immunoblot bands when reacted to an electrophoresed human O type red cell membrane antigen, but those bands had no identity to the cross-reactive bands on the P. falciparum antigen immunoblots. Malaria-positive sera from Papua New Guinean subjects presumed to be uninfected with HIV produced a variety of bands, some of intense prominence, to HIV antigen on diagnostic Western blots. PIP: Serum samples from three populations of Papua New Guinea, where Plasmodium falciparum malaria and human T-lymphotropic virus type 1 (HTLV-1) are coendemic at high prevalence rates, showed statistically significant ELISA co-seropositivity and co-seronegativity. Cross-reactivity was further indicated by the presence of 10 bands ranging from 134 kDa to 18 kDa on immunoblots of electrophoresed whole lysate P. falciparum antigen against serum of HTLV-1 seropositive patients from an area where malaria is not present. Similarly, sera from patients positive for HIV from a nonmalarious region produced immunoblot bands ranging from 134 kDa to 33 kDa to the P. falciparum antigen. The HTLV-1 and HIV serum samples yielded a number of immunoblot bands when reacted to an electrophoresed human O type red cell membrane antigen, but those bands had no identity to the cross-reactive bands on the P. falciparum antigen immunoblots. Malaria-positive sera from Papua New Guinean subjects presumed to be uninfected with HIV produced a variety of bands, some of intense prominence, to HIV antigen on diagnostic Western blots.
Antibodies, Protozoan - immunology ; Enzyme-Linked Immunosorbent Assay ; HIV Seropositivity - immunology ; Papua New Guinea - epidemiology

OBJECTIVETo develop a rapid test kit for antibody to HIV by nano immunomagnetic lateral flow method.

METHODSA rapid test kit was developed by conjugation of the HIV antigen gp41 and gp36 to 200nm super paramagnetic particles by carbodiimide (EDC) and coating of the HIV antigen gp41 and gp36 to nitrocellulose membrane. Then the kit was evaluated with serials of experiments.

RESULTSThe kit was qualified with examination of national reference panel of anti-HIV antibody for colloidal gold diagnostic kit. The sensitivity was 100% by tested with 20 HIV antibody positive sera, the specificity was 98.5% by tested with 600 HIV antibody negative sera, respectively. The stability of the kit was over 12 month by storage at room temperature.

CONCLUSIONA diagnostic kit for antibody to HIV was developed with the advantages of convenience, rapid test, good stability and point of care.

Antibodies, Anti-Idiotypic ; immunology ; Gold Colloid ; chemistry ; HIV ; immunology ; isolation & purification ; HIV Antibodies ; HIV Envelope Protein gp41 ; HIV Infections ; diagnosis ; HIV Seropositivity ; blood ; HIV-1 ; immunology ; isolation & purification ; Immunomagnetic Separation ; methods ; Molecular Biology ; methods ; Nanotechnology ; Reagent Kits, Diagnostic

OBJECTIVETo study the rate of human immunodeficiency virus (HIV) seroconversion, HIV transmission and related risk factors among injecting drug users (IDUs) in an area of Sichuan province.

METHODSIn November 2002, a community-based baseline survey was conducted to recruit 333 HIV-seronegative IDUs for a prospective cohort study in Xichang county of Sichuan province, China. Follow-up visits were carried out every 6 months to study the situation of drug use, sexual behaviors of the IDUs and blood specimens were collected to test for antibodies against HIV and syphilis.

RESULTSDuring a 24-month follow-up period, cohort retention rate and HIV incidence were 75.7% and 2.53 per 100 person-years [95% confidence interval (CI): 1.10-3.97)], respectively. Multivariate Poisson regression model showed that risk factors which were significantly associated with HIV seroconversion would include: ethnicity (RR = 12.42; 95% CI: 2.72-56.74, P = 0.0012) and needle or syringe sharing in the past 3 months (RR = 4.06; 95% CI: 1.29-12.81, P = 0.0168). Syphilis seroconversion in this cohort was 4.71 per 100 person-years (95% CI: 2.59-6.82). In multivariate Poisson regression being female (RR = 4.42; 95% CI: 1.78-10.99, P = 0.0014) appeared to be the only factor which was significantly associated with syphilis seroconversion.

CONCLUSIONOur study results showed that there was a rapid transmission of HIV and syphilis among IDUs in Sichuan province, suggesting that effective intervention should be urgently taken.

Cohort Studies ; HIV Seropositivity ; immunology ; transmission ; Humans ; Prospective Studies ; Risk Factors ; Sexual Behavior ; Substance Abuse, Intravenous ; Syphilis ; immunology ; transmission

OBJECTIVETo explore the mechanisms of malariotherapy for human immunodeficiency virus (HIV)-infected patients and to identify which stage(s) of HIV infection is suitable for the treatment of malariotherapy.

METHODSTherapeutic acute vivax malaria was induced and terminated after 10 fever episodes in 12 HIV-1-infected subjects: Group 1 (G1) had 5 patients with CD(4) T-cell counts >or=500/ micro l at baseline, Group 2 (G2) had 5 patients with CD4 at 499 - 200/ micro l and Group 3 had 2 patients with CD(4) < 200/ micro l (not included in statistical analysis). Enzyme-Linked-Immuno-Sorbent Assay (ELISA) was used to measure plasma levels of cytokines and soluble activation markers. Flow cytometry was used to measure levels of lymphocyte subsets and phenotypes and CD(4) cell apoptosis. Bayer bDNA assay was used to test plasma levels of HIV-1 RNA (viral load). Samples were taken and tested twice before malaria (baselines), three times during malaria and seven times after termination of malaria (at day 10 and 1, 3, 6, 12, 18 and 24 months).

RESULTSLevels of plasma tumor necrosis factor-alpha (TNF-alpha), soluble TNF-alpha receptor-2 (sTNF-RII), neopterin (NPT) and soluble IL-2 receptor (sIL-2R) significantly increased during malaria and sharply reduced to baselines post malaria in all groups. Stronger responses of the aforementioned factors were seen in G2 than in G1 during malaria (P = 0.081, 0.001, 0.013, 0.020). CD4 count and percentage; CD(4)/CD(8) ratio and CD(25)(+) and CD(4)(+)CD(25)(+) percentages increased but HLA-DR+ percentage decreased either during or post malaria in G2. Most G2 patients experienced sustained increase but most G1 patients underwent natural history decline of CD(4) counts and percentages during 2-year follow-up. Percentage of apoptotic CD(4) cells decreased post malaria in all groups. G3 patients had weaker immune responses, however, one advanced AIDS patient in this group experienced clinical improvement after malariotherapy. Most of the 12 patients experienced increase of HIV viral load during malaria but the viral load returned to baseline levels 1 - 3 months after cure of malaria and remained near baseline levels for up to two years.

CONCLUSIONSPart of the mechanisms of malariotherapy is to induce high levels of cytokine activities and subsequently the changes of T-lymphocyte subsets and phenotypes in HIV-infected patients. These findings suggest that malariotherapy may treat HIV-1-infected patients whose CD4 baselines are in the range of 500 - 200/ micro l.

Acute Disease ; Adult ; CD4 Lymphocyte Count ; Cytokines ; blood ; Female ; HIV Seropositivity ; immunology ; therapy ; virology ; HIV-1 ; isolation & purification ; Humans ; Malaria, Vivax ; immunology ; Male ; Viral Load

Based on the computer simulation, we analyzed hydrophobicity, potential epitope of recombined subtypes HIV-1 Env protein (851 amino acids) from Guangxi in China. Compared with conservative peptides of other subtypes in env protein, three sequences (469-511aa, 538-674aa, 700-734aa) were selected to recombine into a chimeric gene that codes three conservative epitope peptides with stronger antigencity, and was constructed in the yeast expression plasmid pPICZB. Chimeric proteins were expressed in Pichia pastoris under the induction of methanol, and were analyzed by SDS-PAGE and Westernblot. The results showed that fusion proteins of three-segment antigen were expressed in Pichia pastoris and that specific protein band at the site of 40kD was target protein, which is interacted with HIV-1 serum. The target proteins were purified by metal Ni-sepharose 4B, and were demonstrated to possess good antigenic specificity from the data of ELISA. This chimeric antigen may be used as research and developed into HIV diagnostic reagents.
Blotting, Western ; Enzyme-Linked Immunosorbent Assay ; Gene Expression Regulation, Viral ; HIV Seronegativity ; immunology ; HIV Seropositivity ; immunology ; HIV-1 ; genetics ; immunology ; metabolism ; Humans ; Pichia ; genetics ; Recombinant Fusion Proteins ; genetics ; immunology ; metabolism ; env Gene Products, Human Immunodeficiency Virus ; genetics ; immunology ; metabolism

To determine laboratory evidence suggesting immunological abnormalities in persons with hemophilia, we evaluated the immunological status of 75 Korean hemophiliacs, seronegative for human immunodeficiency virus (HIV) antibodies, who have been treated only with Korean factor VIII concentrates. From this study, it was shown that Korean hemophiliacs had decreased CD4 levels, increased CD8 levels, and decreased CD4:CD8 ratios. Diminished lymphocyte response to the mitogens, phytohemagglutinin and concanavalin A, and decreased natural killer cell activity were observed in the hemophiliacs. In addition, production of interleukin-II in the hemophiliacs was lower than in the healthy controls. The percentage of B lymphocytes was significantly reduced but the serum levels of immunoglobulin (Ig) G were elevated. However, the serum Ig A and Ig M levels were normal. This study demonstrated a high frequency of immunological abnormalities in HIV antibody negative Korean hemophiliacs treated only with domestic factor VIII concentrates.
Adolescent ; Adult ; Child ; Factor VIII/therapeutic use ; HIV Seropositivity/immunology ; Hemophilia A/drug therapy/*immunology ; Human ; Korea ; Lymphocyte Activation ; Lymphocytes/immunology ; Male ; Middle Age ; Support, Non-U.S. Gov't

OBJECTIVETo study the incidence and risk factors of HIV and syphilis seroconversion among men who have sex with men (MSM) in Beijing.

METHODSA total of 550 MSM were recruited on the basis of community and followed up after 6 and 12 months in Beijing. Each subject was investigated by only one investigator at one time to collect information on demographics and behaviors. Blood samples were collected to test HIV and syphilis seroconversion. ELISA was used for screening test, west blotting (WB) and Particle agglutination were used for confirmatory test.

RESULTSA total of 550 MSM investigated, among which 4.5% (25/550) were HIV-positive and 29.3% (161/550) were syphilis-positive. For 525 HIV-negative MSM, 87.0% (457/525) retained during the 12-month investigation. Seroincidence for HIV and syphilis were 3.37/100 person-years (95%CI = 1.66 - 5.08) and 9.32/100 person-years (95%CI = 5.87 - 12.77) respectively. HIV seroconversions for those who performed and did not perform rectal douching after homosexual anal intercourse in the past 3 months were 7.11/100 and 0.76/100 person-years respectively. Multivariate Cox regression analysis revealed that rectal douching after homosexual anal intercourse in the past 3 months (HR = 9.23, 95%CI = 2.08 - 40.88) was significantly associated with HIV seroconversion. Syphilis seroconversions for those who met male sex partners in parks, public washrooms or bathhouses in the past 3 months were 41.77/100 and 7.97/100 person-years respectively. Syphilis seroconversions for those who performed and did not perform rectal douching after homosexual anal intercourse in the past 3 months were 16.17/100 and 4.92/100 person-years respectively. In the past 3 months, meeting male sex partners in parks, public washrooms or bathhouses (HR = 4.67, 95%CI = 1.77 - 12.34) and performing rectal douching after homosexual anal intercourse (HR = 3.09, 95%CI = 1.40 - 6.83) were significantly associated with syphilis seroconversion.

CONCLUSIONThe seroconversions of HIV and syphilis during the follow-up visits in this MSM cohort study in Beijing were very serious, and that the associated factors for seroconversions were rectal douching after homosexual anal intercourse and meeting male sex partners in parks, public washrooms or bathhouses.

Adolescent ; Adult ; Antibodies, Bacterial ; blood ; China ; epidemiology ; HIV ; immunology ; HIV Antibodies ; blood ; HIV Infections ; blood ; epidemiology ; HIV Seropositivity ; blood ; epidemiology ; Homosexuality, Male ; Humans ; Incidence ; Male ; Risk Factors ; Sexual Behavior ; Syphilis ; blood ; epidemiology ; Treponema pallidum ; immunology ; Young Adult

OBJECTIVETo obtain information on HIV disease progression through western blot (WB) band profile.

METHODS1466 available serum samples of newly reported HIV/AIDS cases, confirmed by WB from Dehong prefecture of Yunnan province in 2011, were tested, using the BED HIV Incidence Capture Enzyme Immunoassay (BED-CEIA) and Limiting Antigen Avidity Enzyme Immunoassay (LAg-Avidity EIA), respectively. Differences of WB band profiled between recent infections and established infections were then compared.

RESULTSInfections defined as 'recent' showed lower mean antibody intensities to p17, p24, p31, gp41, p51, p55, p66, gp120 and gp160 in WB when compared to those established infections with statistical difference(P < 0.05).

CONCLUSIONThese WB band profiles which were considered as 'recent' infections would include factors as:lack of antibody reactivity to gp41 antigen, with intensity scores of all bands lower than 13, and those intensity scores of antibody to env protein lower than 5 or to pol protein lower than 4.

Acquired Immunodeficiency Syndrome ; blood ; diagnosis ; epidemiology ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Blotting, Western ; Child ; Child, Preschool ; China ; Female ; HIV Seropositivity ; HIV-1 ; immunology ; Humans ; Male ; Middle Aged

BACKGROUNDElevated levels of interleukin-7 (IL-7) have been correlated with CD4(+) T cell depletion and the emergence of syncytium-inducing (SI) variants in human immunodeficiency virus type-1 (HIV-1) infection, and suggested as an indicator of acquired immunodeficiency syndrome (AIDS) disease progression. Therefore, we investigated the effects of IL-7 on disease progression and virus phenotype in Chinese HIV/AIDS patients.

METHODSIn a cross-sectional study of 71 untreated HIV-1 seropositive individuals and 12 healthy donors, plasma IL-7 levels were determined by an ultra sensitive enzyme-linked immunosorbent assay (ELISA), and its relations to CD4(+) T cells, CD8(+) T cells, plasma viral loads and HIV phenotypes were analyzed.

RESULTSSignificant higher IL-7 levels were found in Chinese HIV/AIDS patients [(3.33 +/- 3.60) pg/ml] than those of health controls [(1.2 +/- 0.81) pg/ml] (P < 0.05), and IL-7 levels were inversely associated with CD4(+) T cell counts (r = -0.497, P < 0.01). Furthermore, IL-7 levels were significant higher in patients with SI variants [(9.12 +/- 4.55) pg/ml] than those with non-syncytium-inducing variants [(1.50 +/- 2.69) pg/ml] (P < 0.01).

CONCLUSIONSIncreased IL-7 levels were found in Chinese HIV/AIDS patients and significantly associated with disease progression, thus increased IL-7 plasma levels may indicate disease progression.

Adult ; CD4 Lymphocyte Count ; Disease Progression ; HIV Seropositivity ; blood ; immunology ; virology ; HIV-1 ; Humans ; Interleukin-7 ; blood ; physiology ; Phenotype ; Viral Load

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.
False Positive Reactions ; Female ; HIV Antibodies/*blood/immunology ; HIV Core Protein p24/*blood/immunology ; HIV Seropositivity/*diagnosis ; Humans ; Male ; Pregnancy ; Reagent Kits, Diagnostic ; Republic of Korea ; Sensitivity and Specificity ; Time Factors