Triple positivity of HBsAg,anti-HCV antibody,and HIV and their influence on CD4+ lymphocyte levels in the highly HIV infected population of Abeokuta,Nigeria

BackgroundFew studies exist on hospital-based seroprevalence of triple positivity of HIV/HBV/HCV in Nigeria.ObjectivesThe study aimed at determining the triple positivity of HIV, HBsAg and HCV among HIV-infected individuals in Abeokuta, Nigeria and defining the influence of these triple infections on CD4+ counts of HIV-infected individuals as antiretroviral therapy improves in Nigeria.MethodsEnumeration of CD4+ levels in 183 HIV-infected persons was done with Partec Flow Cytometer. Seropositivity of HBsAg and anti-HCV antibody was detected with rapid kits.ResultsFrom the result obtained, significance variance (p<0.05) existed between HIV positive persons and persons who tested positive to HIV/HBV/HCV triple infection before and after the commencement of HAART. Of these infections, 31(16.9%) had HBV/HCV/HIV triple infection, while 152(83.1%) had HIV mono infection only, 56(30.6%) had HBV/HIV dual infection only and 43(23.5%) had HCV/HIV dual infection only. Significant variance (p<0.05) also existed between subjects with CD4 counts of <200 cells/µl, 200–499 cells/µl and >500 cells/µl. Highest seroprevalence of HIV (35.0%) was found in age groups 35–44 years and >65 years had the least (2.7%). Significant variance (p<0.05) also existed in the progression of CD4+ lymphocytes cells between subjects with persistent decrease (32.3%) in CD4+ lymphocytes cells and those with fluctuation in their CD4+ lymphocytes cells (12.9%) after the commencement of ART.ConclusionThe study further confirms that triple positivity of HIV/HBV/HCV infection is common in Abeokuta, Nigeria. Testing of these triple infections should be a big concern in the best choice and commencement of ART. Also, the study showed that consistent and prolonged use of HAART had a positive impact on the CD4 count of HIV-infected individuals.     

Solid-phase radioimmunoassay of serum IgG, IgM, and IgA antibodies to cytomegalovirus

A radioimmunoassay (RIA) using polystyrene beads as the solid phase for cytomegalovirus (CMV) antigen and iodinated immunosorbent purified anti-human IgG, IgM, and IgA as indicator antibodies was developed for the detection of immunoglobulin class-specific antibodies to CMV. An antigen prepared from extracellular virus was essential for reliable results, and a preparation ultracentrifuged and sonicated twice was better than a crude antigen. The optimal antigen gave low cpm values with a negative reference serum, resulting in cpm ratios of 10 or higher between early convalescent phase serum and negative reference serum. Of six patients with an increase in CMV CF titres, all six had an increase in RIA IgG titres, four had an increase in IgA titres, and all had IgM antibodies. The IgG titres were high, up to 1/64,000. In a group of 17 infants negative in CMV CF test, 14 had CMV IgG antibodies in RIA test, indicating mainly low levels of maternal antibodies. In six of seven patients with CMV isolations from urine specimens, an increase in IgG or IgA titres or the presence of IgM antibodies was found, and only one of these patients had an increase in CMV CF titre. The specificity of the developed CMV RIA test was further demonstrated by detecting no significant increase in RIA titres in serum specimens of patients with primary herpes simplex infection, chickenpox, herpes zoster, or infectious mononucleosis.     

Solid-phase radioimmunoassay of IgA, IgG, and IgM antibodies to human rotavirus.

A solid-phase radioimmunoassay (RIA) has been developed for the detection of human rotavirus-specific IgA, IgG, and IgM antibodies. Nebraska calf diarrhea virus grown in LLC-MK2 cell cultures in the presence of trypsin was directly adsorbed onto polystyrene balls, and antibodies that attached to the virus-coated balls were detected by subsequent binding of 125I-labeled antibodies specific to human alpha, gamma or mu chains of human Iga, IgG, or IgM immunoglobulins. A total of 116 serum specimens from 58 adult patients were tested. Binding ratios between the positive and the negative serum varied between 5 and 15, occasionally being 20 or more in the IgA and IgG assays, but rarely exceeding 3 in the IgM assay. The RIA was found to be more sensitive in detecting antibodies to rotavirus than the complement fixation (CF) test, the RIA titers obtained being 50--100 times as high as the CF titers. The method described offers a possibility of evaluating the immune response to human rotavirus and of detecting recent infection.     

Reverse radioimmunoassays of IgM and IgG antibodies to Coxsackie B viruses in patients with acute myopericarditis

A reverse radioimmunoassay (RIA) of antibodies to enteroviruses, previously developed for the detection of IgM antibodies to Coxsackie B1 (CB1) and B3 (CB3) and to Echo 11 (E11) and 30 (E30) viruses, was extended in the present study for the detection of IgM antibodies to Coxsackie B2 (CB2), B4 (CB4), and B5 (CB5) viruses and of IgG antibodies to CB1-CB5, E11, and E30 viruses. After standardisation of the assays and application to a collection of serum specimens from patients with proven enterovirus infections, specimens from patients with diagnosed or suspected acute myo- and/or pericarditis (myopericarditis group), and control specimens from patients with nonenterovirus infections, were studied, as well as from apparently healthy subjects. Of the patients with enterovirus infections, 29 of 30 (97%) were positive in the IgM RIA and 19 of 25 (76%) in the IgG RIA. In the myopericarditis group, 18 of 37 (49%) patients showed Coxsackie B (CB) virus-specific IgM titres and 9 of 37 (24%) CB virus-specific IgG titres. In the control specimens very few positive responses were detected. The RIAs appeared to be type specific or at least predominantly type specific, provided that the amount of labeled virus was carefully standardised. The sensitivity of the RIAs seemed to be rather high for IgM but low for IgG. In the neutralisation (NT) test no significant rise or fall in titre against CB viruses was demonstrated in the myopericarditis group. It is concluded that the reverse IgM RIA may be valuable for studies of the role of CB viruses in acute myo- and/or pericarditis.     

Cytomegalovirus (CMV) infection, CD4+ lymphocyte counts and the development of AIDS in HIV-1-infected haemophiliac patients.

After a maximum of 11 years (median 8.3 years) from the time of HIV seroconversion, 25 out of 59 (42%) of CMV-seropositive haemophiliacs had progressed to AIDS, as opposed to eight out of 50 (16%) CMV seronegatives. The age-adjusted relative risk for AIDS among CMV seropositives was 2.4 (P = 0.03). In order to determine how this adverse effect is mediated, the mean rate of decline in serial CD4+ lymphocyte counts was studied. CD4+ lymphocyte counts tended to decline more rapidly in CMV seropositives than in seronegatives (-0.087 x 10(9)/l per annum versus -0.082 x 10(9)/l per annum), but this difference did not reach statistical significance. The average CD4+ lymphocyte count at the time of HIV seroconversion was estimated to be similar in CMV seropositives and negatives, because in HIV-1-negative haemophiliacs the CD4+ counts were virtually identical, after adjustment for age (0.94 x 10(9)/l and 0.97 x 10(9)/l, respectively). The median CD4+ cell count at which AIDS developed was higher in the CMV-seropositive group (0.07 x 10(9)/l) than in the seronegative group (0.04 x 10(9)/l), but this difference did not reach statistical significance. We conclude from these findings that the adverse effect of CMV is not wholly mediated via a more rapid loss of CD4+ cells. We discuss other processes that may be mediated by CMV, such as a functional deficiency of residual CD4+ cells, or dissemination of HIV in other organs, which may be important in determining the earlier onset of AIDS among CMV-seropositive subjects.     

Determination of human IgG and IgM class antibodies to West Nile virus by enzyme linked immunosorbent assay (ELISA)

An enzyme-linked immunosorbent assay (ELISA) was developed and used for the detection of IgG and IgM antibodies to West Nile virus in human sera. Thirteen paired sera of clinical cases and 24 control sera taken randomly from a blood bank repository were tested. The sera were reacted in microtiter plates coated with PEG-treated WNV antigen. IgG or IgM antibodies were quantitated by the use of alkaline-phosphatase-conjugated anti-human IgG or IgM antibodies. Of the 24 randomly collected serum samples, 7 were positive in the IgG-ELISA test. One positive by the IgM-ELISA was found to contain rheumatoid factor. In 12 of 13 paired sera of clinical cases, IgM as well as IgG antibodies were detected in the second serum sample taken about 3 wk after the onset of clinical signs. The IgM positive sera were screened for rheumatoid factor (RF) on IgG-coated plates. None of them contained RF. Antibody titers obtained by ELISA showed a good correlation with titers obtained by hemagglutination inhibition, complement fixation, and neutralization tests. The ELISA tests for detection of IgM and IgG antibodies to WNV therefore can replace the other serological methods for epidemiological surveillance and diagnostic purposes.     

Chronological evolution of IgM, IgA, IgG and neutralisation antibodies after infection with SARS-associated coronavirus

Abstract Serum levels of IgG, IgM and IgA against severe acute respiratory distress syndrome (SARS)-associated coronavirus (SARS-CoV) were detected serially with the use of immunofluorescent antibody assays in 30 patients with SARS. Seroconversion for IgG (mean 10 days) occurred simultaneously, or 1 day earlier, than that for IgM and IgA (mean 11 days for both). IgG could be detected as early as 4 days after the onset of illness. The earliest time at which these three antibodies reached peak levels was similar (mean 15 days). A high IgG level (1:800) could persist for > 3 months. The kinetics of neutralisation antibodies obtained with 100x the tissue culture infective dose (TCID50) of the SARS-CoV TW1 strain in five patients with SARS nearly paralleled those for IgG. There were no significant differences in the kinetics of the IgG, IgM and IgA responses between patients with or without underlying medical disease, steroid or intravenous immunoglobulin therapy, or mechanical ventilation.     

Detection of Human Papilloma Virus (Type 16) among HIV-Positive Women in Ogbomoso,South-Western Nigeria

Background: Human papilloma virus (HPV) is a common sexually transmitted virus which infects the cutaneous and mucosal epithelium. HPV Type 16 is one of the viruses that causes cervical cancer and immunocompromised individuals are at high risk of different co-infections. Women living with Human Immunodeficiency Virus (HIV) have greater risk to the virus due to their impaired immunity. This study aimed at determining the seroprevalence of HPV IgM (Type 16) among HIV-infected women in Ogbomoso.

Methods: The blood sample of 180 consenting subjects were obtained and their sera subjected to serological assay using Enzyme Linked Immunosorbent Assay. Samples were collected over a period of 6 months (July–December 2014).

Results: The mean age and mean CD4+ count of the subjects was 38.22 ± 0.79 years and 392.80 ± 20.98 cells/μL, respectively. Out of 180 subjects tested, 18 (10%; 95% confidence interval) were positive for HPV Type 16 IgM. HPV Type 16 IgM was highest among the age group 31–45 (61.11%), traders (38.89%), >500 CD4/μL (33.33%). The seroprevalence using logistic regression at P < 0.05 shows there is a significant difference between the age and CD4 ⁺ cell count.

Conclusion: The result provides evidence that HPV Type 16 is present among HIV-infected women in Ogbomoso and they are susceptible to cervical cancer. This seroepidemiological survey is important for the prevention efforts such as availability of vaccine.     

IgG subclass reactivity against human immunodeficiency virus (HIV) and cytomegalovirus in cerebrospinal fluid and serum from HIV-infected patients

Cerebrospinal fluid (CSF) and serum samples from 17 patients seropositive for the human immunodeficiency virus (HIV) were analysed for specific IgG1-4 against HIV and cytomegalovirus (CMV). Measles IgG was studied as a reference to detect blood-brain barrier (BBB) defects. All patients had IgG1 antibodies against HIV in both CSF and serum, and all had CMV IgG1 in serum (16 in CSF). Anti-HIV IgG was synthesised intrathecally in 11 patients, IgG3 in three patients, and IgG4 in three patients. Intrathecal production of anti-CMV IgG1 was found in three patients, IgG2 in one, IgG3 in three, and IgG4 in one. Intrathecal anti-HIV IgG synthesis could be demonstrated in all stages of the disease. Analysis of all IgG subclasses allowed intrathecal HIV and/or IgG production to be detected also in patients in whom intrathecally synthesised IgG was restricted to IgG2, 3, or 4. The expression of HIV-specific IgG subclasses in CSF and serum was more restricted in AIDS patients than in HIV-infected persons without clinical AIDS. On the contrary, the largest number of CMV-specific IgG subclasses was found in AIDS patients. Intrathecal HIV or CMV IgG subclass production was seen both with and without neurological symptoms. The peripheral T4 cell counts were not obviously related to neurological symptoms. Even patients with low peripheral T4 cell counts had evidence of intrathecal antibody synthesis against HIV and sometimes CMV, suggesting a retained helper function of T cells in the central nervous system.     

高效抗病毒治疗促使艾滋病患者免疫功能重建

艾滋病的特征是ＨＩＶ 1感染人体后 ,造成ＣＤ4 +Ｔ淋巴细胞数量进行性减少、细胞免疫功能损害 ,最后导致艾滋病 (ＡＩＤＳ)。先前的研究表明这种免疫功能的丧失是不可逆转的 ,抗ＨＩＶ病毒治疗仅能控制或减缓其进展。近年来 ,由于强效联合抗病毒治疗 (ＨＡＡＲＴ)的应用 ,艾滋病的发病率和死亡率均较前明显下降 (指西方国家 )。说明ＨＡＡＲＴ不仅能有效的控制ＨＩＶ 1的复制 ,并能使艾滋病病人的免疫功能得到恢复。这种ＨＡＡＲＴ使艾滋病病人免疫功能重建的假说最近被一组法国研究人员证实 ,艾滋病的免疫重建规律是 :(1)治疗早期ＣＤ4 +…     

Interleukin-2 reconstitutes defective human immunodeficiency virus (HIV), and cytomegalovirus (CMV) specific CD8+ T cell proliferation in HIV infection

Recent studies indicate that a defective proliferative response of HIV-specific CD8+ T cells is associated with the lack of virologic control in chronic HIV infection in humans. The possible mechanisms that might be responsible for the reduced proliferative potential of HIV-specific CD8+ T cells and conditions conducive to the proliferation of CD8+ T cells were examined in 14 HIV-infected individuals and 7 HIV-uninfected controls using CFSE labeling and flow cytometry techniques, and analyzed data using 2 quantitative measurements: the percentages of proliferating CD8+ T cells (Tp), and the maximum number of cell divisions (Dm) after stimulation. It was found that CD8+ T cells from HIV-infected and -uninfected subjects proliferated equally well after polyclonal stimulation by phylohemagglutinin A (PHA); both groups reached a Tp of 92%-96% and a Dm of 5-8. However, in HIV-infected subjects, proliferation of HIV- and CMV-specific CD8+ T cells was significantly reduced compared to proliferation of CMV- specific CD8+ T cells from HIV-uninfected subjects. These defective proliferative responses of HIV- and CMV-specific CD8+ T cells were restored by the addition of IL-2 at the time of stimulation. These results may have implications for the design of immune modulation strategies in vivo.     

人体包虫病主要特异性免疫球蛋白的水平及其临床意义

本文报道了应用酶联免疫吸附试验(ELISA)检测包虫病人血清中抗包虫特异性免疫球蛋白(IgG和IgM)的水平并对其临床意义进行了初步探讨。48例正常人血清特异性IgG和IgM的假阳性检出串均为0％;24例囊尾蚴病人血清特异性抗包虫IgG和IgM的假阳性检出效分别为:33.4％;4.16％;42例包虫病患者血清特异性IgG和IgM的阳性检出率分别为:80.95％,66.67％;两者之差经统计学处理具有显著意义(p<0.05)。肝包虫特异性IgG的阳性检出率较肺包虫高,肺包虫特异性IgM的阳性检出率较肝包虫高。这提示患者对包虫的免疫应答和其寄生的位置有关;诱导产生的特异性免疫球蛋白主要为IgG类,特异性IgM的水平与包虫囊壁的完整与否以及检测方法的灵敏性、特异性有关。     

A modified passive-haemagglutination technique for the detection of cytomegalovirus and herpes simplex virus antibodies: application in virus-specific IgM diagnosis

Cytomegalovirus (CMV) and herpes simplex virus (HSV) antibodies were detected by a modified passive haemagglutination (PHA) technique. The main features of this modification are the use of a simpler method for the removal of nonspecific sheep agglutinins in the sera, the deployment of commercially available CMV and HSV antigens, and a different sucrose density gradient (SDG) system for the separation of IgM from IgG. The modified procedure proved to be a reliable, specific, and sensitive technique in detecting antibodies to CMV and HSV in both whole serum and in the separated IgM and IgG fractions. It was as reliable as the complement-fixation (CF) test when applied in seroepidemiological studies and in the detection of antibodies in cord serum. Preliminary data are provided which suggest that the combination of SDG and PHA may prove to be a more reliable system for the detection of exclusion of CMV-specific IgM than an enzyme-linked immunosorbent assay (ELISA).     

抗CD4及抗CXCR4抗体阻断HIV—1感染细胞作用的研究

为探讨抗ＣＤ４ 及抗ＣＸＣＲ４ 抗体在阻断Ｉ型人免疫缺陷病毒（ＨＩＶ－ １） 感染应用中的意义, 本文应用上述两种抗体分别与ＳｕｐＴ１ 细胞及人外周血单个核细胞（ＰＢＭＣ） 共培育, 以封闭ＨＩＶ－ １ 在上述细胞上的受体。然后, 以ＨＩＶ１ ＮＬ４３ 病毒株感染上述细胞, 通过测定感染细胞上清中ＨＩＶ－ １ 的Ｐ２４ 蛋白含量, 观察上述抗体对ＨＩＶ－ １ 感染细胞的阻断作用。结果显示, 无论是抗ＣＤ４ 或抗ＣＸＣＲ４ 的抗体单独应用或是两者联合应用, 均可明显地抑制ＨＩＶ－ １ 感染细胞的作用。该结果为今后开拓ＡＩＤＳ的抗体治疗提供了理论基础。     

An enzyme labelled nuclear antigen immunoassay for detection of cytomegalovirus IgM antibodies in human serum: specific and non-specific reactions

A mu-capture enzyme linked immunosorbent assay was developed for detection of IgM antibody to cytomegalovirus (CMV). Virus-specific IgM was detected using horseradish peroxidase labelled nuclear CMV antigen (CMV-ELA). False-positive reactions caused by Paul-Bunnell-Davidsohn (PBD) positive sera and antinuclear antibody (ANA) positive sera were identified in a combination assay employing enzyme labelled nuclear control antigen (CO-ELA) in parallel to the CMV-ELA. Four of five PBD positive and 30 of 31 ANA positive sera reactive with the CMV-ELA were identified as false positive reactions in the combined ELA-assay. The reactivity in PBD-positive sera could not be explained by antigenic cross reactivity between CMV and Epstein-Barr virus, and the results further suggested that different cell specified components of the CMV-ELA were responsible for the reactivity of PBD-positive as compared to ANA-positive sera. One of 314 healthy blood donors, 12 of 12 patients with primary CMV infection, and 11 of 15 patients with secondary CMV infection had detectable CMV IgM antibodies. Comparison of different CMV-ELAs revealed that pronounced differences in specificity as well as sensitivity may exist.     

Specific recognition pattern of IgM and IgG antibodies produced in the course of experimental paracoccidioidomycosis.

Specific IgM and IgG responses to Paracoccidioides brasiliensis produced in resistant and susceptible mice during experimental paracoccidioidomycosis were examined by the immunoblotting procedure. Sera from infected mice recognized 51 antigen bands with apparent molecular masses from 8 to 86 kD. Sixteen of these were defined as major antigen bands because of almost universal presence of antibodies to them, and their intense staining. All sera, including those from normal control mice, tested for both IgM and IgG antibody reacted with the major E antigen which appeared as a large diffuse band from 43 to 47 kD. Comparisons between resistant and susceptible mice showed some significant differences in IgM responses to many antigen bands. While IgG responses were quite similar for both strains, differences were apparent in the response to the antigens at 62 and 68 kD.     

HIV/AIDS患者高效抗逆转录病毒治疗后免疫重建不良的特征及相关因素分析

目的 通过回顾性研究,描述免疫重建不良HIV/AIDS患者的基本特征,探讨免疫重建不良的相关因素,发现HIV/AIDS患者免疫重建不良的预示指标.方法 以2005年1月-2011年8月在北京地坛医院起始HAART治疗≥96w的HIV/AIDS患者（503例）为研究对象,比较其中免疫无应答者（76例）与免疫应答良好者（427例）临床资料（人口学资料、基线CD4＋T细胞计数、机会性感染的合并情况、HAART（高效抗逆转录病毒治疗）方案和各种化验指标的差异,分析HIV/AIDS患者免疫重建不良的相关因素.结果 入组病例中免疫重建不良的发生率为15.11％;免疫重建不良组的基线CD4＋T细胞数、总淋巴细胞数目（TLC）及血红蛋白（Hb）显著低于免疫重建良好组（z=-9.056,P＜0.001;z=-5.541,P＜0.001;z=-3.014,P=0.003）.免疫重建不良组基线机会性感染合并率显著高于重建良好组（x2=14.834,P=0.037）.结论 低基线CD4＋T细胞数目、TLC及Hb均为免疫重建不良的相关因素,应对HIV感染的患者及早进行治疗,预防免疫重建不良的发生.     

Cytomegalovirus IgG and IgA serum antibodies in a study of HIV infection and HIV related diseases in homosexual men.

The significance of serum IgG and IgA antibodies to cytomegalovirus (CMV) at various stages of human immune deficiency virus (HIV) infection was studied in 175 homosexual men. Sera were obtained from 123 HIV seropositives [41 asymptomatic, 29 with lymphadenopathy associated syndrome (LAS), 22 with AIDS related complex (ARC), and 31 AIDS patients], 17 HIV seroconverters, and 35 HIV asymptomatic seronegatives. The sera were tested blindly for CMV IgA and IgG antibodies using the immunoperoxidase assay (IPA) and CMV infected human embryo cells. Cross-sectional analysis of CMV IgG antibodies at a titer of greater than or equal to 20 showed 87% and 100% prevalence in the HIV seronegative groups and in the HIV seropositive groups, respectively (P less than 0.05). CMV IgG antibodies at a titer of greater than or equal to 80 were present in significantly higher proportions among the HIV seropositive subjects of the various groups as compared with the HIV seronegative homosexual men. However, in the HIV seronegatives who later seroconverted to HIV, a significantly higher prevalence of CMV antibodies (35%) was detected before HIV seroconversion, as compared with the persistently HIV seronegative subjects (14.3%) (P less than 0.05). The HIV seronegatives pre-HIV seroconversion also exhibited a significantly higher geometric mean titer (GMT) of CMV IgG antibodies (62.17 +/- 0.64) as compared with the persistently HIV seronegatives (34.0 +/- 0.6) (P = 0.03). Significantly higher GMTs of CMV IgG antibodies were detected in all the HIV seropositive groups as compared with the persistently HIV seronegative group. CMV IgG antibodies were not detected in the HIV seronegative subjects.(ABSTRACT TRUNCATED AT 250 WORDS)     

Evaluation of the Abbott AxSYM cytomegalovirus (CMV) immunoglobulin M (IgM) assay in conjunction with other CMV IgM tests and a CMV IgG avidity assay

The measurement of the avidity of cytomegalovirus (CMV) immunoglobulin G (IgG) antibodies has been shown by several investigators to be useful in identifying and excluding primary CMV infections in pregnant women. In this work, we examined the diagnostic utility of reflex testing of CMV IgM-positive specimens from pregnant women by using a CMV IgG avidity assay. The utility of this approach was directly dependent on the sensitivity of the CMV IgM assay employed during the initial screen. The higher initial reactivity rate of the AxSYM CMV IgM assay was necessary in order to detect CMV IgM in specimens containing low-avidity CMV IgG antibodies, indicative of a primary CMV infection, which other CMV IgM assays (Behring, Vidas, Captia, and Eurogenetics) fail to detect in some cases. The use of the AxSYM CMV IgM assay, followed by an avidity test, should result in more accurate diagnosis of CMV infection in pregnant women.