Immunological study of cytomegalovirus complement-fixing antibodies in the population of the CSR.

The population from different regions of the CSR was examined serologically for the presence of cytomegalovirus (CMV) antibodies. Exclusively persons showing no signs of disease, chosen by random selection, were examined. The immunological survey was carried out by the complement fixation reaction with CMV antigen prepared in the laboratory from the international strain AD 169. The high incidence found in normal population is suggestive of a considerable dispersion of cytomegalovirus infection. The contact with the virus is followed by antibody response, manifested in most cases only subclinically and asymptomatically. The incidence of CMV antibodies increases with increasing age. In young age groups, antibodies were found in approximately 20%. The number of persons showing positive reactions increased gradually and in the oldest age groups antibodies were found in 70%. The significance of factors causing the incidence of cytomegalovirus antibodies in the population is discussed.     

Seroepidemiologic study of herpes simplex virus type 2 and cytomegalovirus among young adults in northern Jordan

Blood samples were randomly collected from 360 males and 390 females among apparently healthy university students aged 18-24 years and tested for herpes virus type 2 (HSV-2) and cytomegalovirus (CMV) antibodies. The prevalence of HSV-2 seropositivity was 52.8% for males and 41.5% for females as detected by ELISA. On the other hand, the prevalence of CMV seropositivity was 75.6% in males and 77.2% in females. The high percentage of seropositivity in our study is most probably due to the crowded living and low socioeconomic status of the Jordan population. The higher prevalence in males could be due to the way of life in the Middle East that gives males the freedom to play outdoors more than females.     

Serological markers of hepatitis B virus and cytomegalovirus infections in Norwegians with coagulation factor defects

The prevalence of serological markers for present and past hepatitis B virus (HBV) infection and antibodies against cytomegalovirus (CMV) among Norwegians with coagulation factor defects was examined in serum samples collected before virus-inactivated coagulation concentrates came into use. Sera collected in 1985/86 from 324 of 377 (86%) registered persons with such defects were available. Three persons were chronic carriers of HBsAg. The prevalence of HBV antibodies was 28% compared with about 5% in the general population. The highest prevalence rate was found among patients with severe haemophilia A (44%) and in patients with haemophilia B (39%). The prevalence of anti-CMV antibodies was 75% which is similar to that found in the general Norwegian population.     

Prevalence of Cytomegalovirus Infection in a Normal Urban Population in Nova Scotia

During 1967 and 1968 a seroepidemiological survey was conducted on the prevalence of CF antibodies to CMV in the normal population of the Halifax-Dartmouth metropolitan area of Nova Scotia. Samples of 550 sera, collected from all age groups, including newborns, were processed. At birth, 34% of infants possess antibodies, presumably of maternal origin, and there follows a decline until at 2 years of age only 4% of children exhibit evidence of infection. There is a gradual increase up to 16% by 20 years of age, and thereafter this is maintained until by 40+ years half the population possesses antibodies.A more detailed analysis of cord sera indicated that approximately 60% of women of childbearing age possess no antibodies and are susceptible to primary CMV infection. Among these, age and gravidity are not significant factors.These data indicate that CMV infection is fairly widespread in this community, although comparative studies suggest that it is less so than in some other areas, such as Easter Island, where a more homogeneous pattern of overcrowding and poor sacioeconomic conditions prevails.     

Prevalence of cytomegalovirus infection and its role in total immunoglobulin pattern in Iranian patients with different subtypes of multiple sclerosis

Multiple sclerosis (MS) is the most common autoimmune disease characterized by multifocal areas of inflammatory demyelination within the central nervous system. Cytomegalovirus (CMV) has a complex pathobiology and in most cases is simply asymptomatic. There is some recent controversy over the role of CMV in the pathology of MS. The aim of this study was to evaluate active CMV infection and its effect on the humoral immunity in patients with MS. Serum, plasma, peripheral blood mononuclear cells (PBMCs), saliva and urine collected from MS patients (n=78) and healthy subjects (n=123) were screened for the presence of anti-CMV antibodies and CMV-DNA by nephelometric and PCR methods. Concentrations of total antibodies in MS subtypes were measured using both nephelometric and enzyme linked fluorescent assay (ELFA) techniques. The results extend the observation of an increased frequency of CMV-DNA in patients, in contrast with controls (p<0.001). Furthermore, systemic CMV infections were found in 25.5% of patients and only 3.2% of controls (p<0.001). There was significant difference in the titers of anti-CMV IgG and total IgE in patient and controls (P<0.001). These results support the hypothesis that CMV may contribute to MS thought to establish systemic infection process and induce immune response.     

Associations between HLA-DQB1 high-risk alleles and type I diabetes do not depend on cytomegalovirus antibody status at onset: a case-parent study conducted in Chile

The purpose of the present study is to ascertain whether the associations between HLA-DQB1*0201 and DQB1*0302 alleles and childhood diabetes depend on the presence of antibodies to human cytomegalovirus (CMV). A study of incident type I diabetes cases and parents was conducted in Santiago, Chile. HLA-DQB1 polymorphisms were determined in 85 case-parent trios (255 subjects), while the detection of CMV was carried out only in the incident cases. As expected, HLA-DQB1 polymorphisms are strongly associated with type I diabetes, with crude odds ratios of 3.7 (95% confidence interval (CI) 1.8-7.7) for the DQB1*0201 allele and 10.3 (95% CI 5.0-21.4) for the DQB1*0302 allele. In the subset of families with CMV+ cases, the odds ratios were estimated as 3.7 (95% CI 1.6-8.6) for the DQB1*0201 allele and 11.1 (95% CI 4.8-25.8) for the DQB1*0302 allele. In families with patients who tested negative for CMV antibodies, the odds ratios were calculated as 3.5 (95% CI 0.7-16.8) for the DQB1*0201 allele, and 8.0 (95% CI 1.8-34.7) for the DQB1*0302 allele. There was no evidence of statistical interaction between CMV antibodies and the DQB1*0201 allele (P value = 0.9) or the DQB1*0302 allele (P value = 0.7). In conclusion, alleles DQB1*0302 and DQB1*0201 do not display distinct associations with type I diabetes depending on the presence of antibodies for CMV.     

Comparative efficacy of laboratory methods for cytomegalovirus detection in autopsy material

Eighty eight autopsy specimens obtained from 30 fetuses, still-borns and infants died during the first year of life, all suspected for congenital virus infection at postmortem examination, were studied. The specimens were analyzed by 3 techniques: rapid culture method (RCM) for detection of cytomegalovirus (CMV) infectious activity, the immunocytochemical method for detection of CMV antigen in prints of organs and polymerase chain reaction (PCR) for detection CMV DNA. CMV was detected in 16 out of 26 specimens (61.5%) by PCR, in 43 out of 88 specimens (49%) by RCM and in 15 out of 64 specimens (23%) in prints. The comparison of immune reagents revealed that monoclonal antibodies (McAb) were more specific than polyclonal serum antibodies, as the latter yielded the positive reaction in 10 out of 26 cases (38%), found to be negative in PCR. The data thus obtained indicate that complex techniques, including PCR and RCM in combination with McAb, should be used for evaluation of CMV infection role in child mortality.     

The role of IgG avidity in diagnosis of cytomegalovirus infection in newborns and infants

To evaluate the value of IgG avidity in diagnosis of congenital cytomegalovirus (CMV) infection in newborns and infants we collected serum samples from 40 infants under 12 months of age with suspected congenital CMV infection. Sera were tested for IgM, IgG and IgG avidity. For 25 of them, virus isolation and/or polymerase chain reaction (PCR) on urine specimens were performed. Thirteen (32.5%) patients showed the presence of CMV IgM antibodies, 3 (7.5%) had equivocal IgM result, and 24 (60.0%) patients had IgG antibodies only. Using IgG avidity, CMV infection (low avidity index-AI) was documented in 61.5% IgM positive and 54.2% IgM negative patients. Eight of nine (88.8%) IgM positive patients were positive either on virus isolation or PCR. In IgM negative patients, 46.6% urine cultures were positive for CMV and 66.6% were PCR positive. According to age, IgG avidity demonstrated acute/recent primary CMV infection in 58.8% patients younger than three months compared with 91.7% and 81.8% in 3-6 and 6-12 months old babies, respectively. In conclusion, IgG avidity is useful in diagnosis of CMV infection either in IgM positive or IgM negative children older than 3 months of age. In infants less than 3 months, transplacentally derived maternal IgG antibodies of high avidity influence on the IgG avidity result. In these children, CMV infection should be confirmed by direct virologic methods such as virus isolation or PCR.     

Enzyme linked immunosorbent assay (ELISA) for determination of IgG antibodies to human cytomegalovirus

A solid-phase enzyme linked immunosorbent assay (ELISA) for determination of IgG antibodies to cytomegalovirus (CMV) is described. The assay used purified CMV and extracts of CMV infected cells as antigen. Antigens were desiccated onto the bottom surface of polystyrene microcuvettes. The antibodies bound to the antigens were assayed by anti-IgG-alkaline phosphate conjugate followed by addition of the enzyme substrate. Titration curves have been obtained from the sera of 35 blood donors and of 23 patients. Comparison of results obtained by ELISA with those obtained by complement fixation (CF) shows that there is agreement between the tests. Both purified CMV and extracts of CMV infected cells were found to be suitable antigens. Purified CMV was of value particularly in those sera which show high reactivity against control antigen. The ELISA technique described is approximately 412 to 548 times more sensitive than the CF test when purified CMV or extracts of CMV infected cells, respectively, are used as antigens. No significant heterotypic rise to CMV was observed by ELISA in three sets of sera with seroconversion to herpes simplex virus. The ELISA technique gives objective results, is easily performed, and may be adaptable as a routine test both for serological diagnosis of CMV infection and for screening of the general population.     

Epstein-Barr virus and cytomegalovirus antibodies in infectious mononucleosis.

A method has been evolved for the demonstration of Epstein-Barr virus (EBV) and cytomegalovirus (CMV) infection in 83 cases of infectious mononucleosis. Serum samples were tested for EBV IgM, anti-VCA IgG, anti-EBNA, CMV IgM and CMV IgG antibodies. An acute-phase sample (or samples) and a convalescence sample were examined in each case, and in 44 cases an additional samples was examined 5-12 months after the illness. Since the different antibodies showed characteristic differences in both titre and persistence, a reliable serodiagnosis has become possible. Acute EBV infection is characterized by the presence of EBV-VCA IgG and EBV IgG antibodies and the lack of anti-EBNA. The latter becomes demonstrable as late as the 4th to 5th month after infection. Mean age of the patients was 19 years. EBV infection was demonstrated in 65%, CMV infection in 18% of the cases. In 12% double infection seemed to be probable.     

Rapid diagnosis of cytomegalovirus in immunocompromised patients using monoclonal antibodies that detect early viral antigens

A technique was applied to detect early fluorescent antigens (DEFA) of cytomegalovirus (CMV) using the E13 monoclonal antibodies in 52 immuno-compromised patients hospitalized in the Nephrology Institute of Havana. Of the 75 urine or blood (buffy coat) samples taken, 15 were found positive to CMV. Using classical diploid human fibroblast isolation technique, 12 CMV strains were isolation of previously detected positive samples by DEFA. In addition, CMV was isolated from one sample reported to be negative by DEFA. A coincidence of 80% was found between both techniques. With the ELISA test, all the sample studied have IgG antibodies to CMV.     

Cytomegalovirus infection in Omsk region

Epidemiologic studies of prevalence of cytomegalovirus infection (CMVI) in Omsk region during a period of 15 years showed 1.6-fold increase of population seropositivity to CMV with rate of increase +2.94. In recent years antibodies to CMV were detected in 90.3 - 94.8% of cases. Diagnostics of innate and acquired CMVI was improved. Feasibility of detection of infectious process activity during complex use of new laboratory methods (IgA detection, assessment of IgG avidity, IgG to individual antigens in immunoblotting as well as detection of DNA and "early phase" viral ptoteins) was demonstrated. High rates of infection in population determine the necessity of revision of strategy for examination of children and adults, including screening of pregnant woman, on this infection.     

Detection of the markers of herpes simplex virus and cytomegalovirus in newborns and infants

A total of 111 children suspected for herpesvirus infection were examined. In blood and urine samples the infectious activity of herpes simplex virus (HSV) and cytomegalovirus (CMV) was detected by the rapid culture method (RCM) and the presence of virus DNA--by the polymerase chain reaction (PCR). HSV and/or CMV were detected by two laboratory methods in 57 examined children (51%). Of these, in 18 children (16.2%) both HSV and CMV were detected. The coincidence of the results of the detection of HSV and CMV by these two methods was observed in 72.4% and 75.2% of cases respectively. The comparative analysis of the detection of anti-CMV IgG and IgM was made with the use of commercial test systems produced bythe following manufacturers: "Vector-Best" and "Bioservice" (Russia), "HUMAN" and "Boehringer" (Germany). The effective detection of both anti-CMV (IgG and IgM) was ensured by the test systems "Boehringer". The test system "Vector-Best" for anti-CMV IgG proved to be not inferior as regards sensitivity and specificity. The German test systems demonstrated the highest specificity in the detection of low-avid antibodies. The data obtained in this study indicate that the detection rate of HSV and CMV markers in newborns and infants suspected for herpesvirus infection was, on the average, 20 - 40%. Reliable diagnostics in newborns and infants is possible only in the presence of the combination of at least 2 serological tests (the determination of antivirus IgM and IgG avidity) and 2 methods for the detection of direct herpesvirus markers (PCR and RCM).     

Humoral and cellular CMV responses in healthy donors; identification of a frequent population of CMV-specific, CD4+ T cells in seronegative donors

CMV status is an important risk factor in immune compromised patients. In hematopoeitic cell transplantations (HCT), both donor and recipient are tested routinely for CMV status by serological assays; however, one might argue that it might also be of relevance to examine CMV status by cellular (i.e., T lymphocyte) assays. Here, we have analyzed the CMV status of 100 healthy blood bank donors using both serology and cellular assays. About half (56%) were found to be CMV seropositive, and they all mounted strong CD8+ and/or moderate CD4+ T cell responses ex vivo against the immunodominant CMV protein, pp65. Of the 44 seronegative donors, only five (11%) mounted ex vivo T cell responses; surprisingly, 33 (75%) mounted strong CD4+ T cell responses after a brief in vitro peptide stimulation culture. This may have significant implications for the analysis and selection of HCT donors.     

Association between antibody response against cytomegalovirus strain-specific glycoprotein H epitopes and HLA-DR

The gH of CMV is a major target for strain-specific neutralizing antibodies. To verify whether there is a correlation between HLA-DR type and strain-specific antibodies, antibodies against CMV gH in potential donors and recipients for renal transplantation were investigated. Among 471 subjects, 404 (86%) showed reactivity to CMV gH, but no antibodies against gH were detected in 67 (14%) subjects. The positive rates were over 80% in most HLA subpopulations. Fewer subjects with HLA-DR10 and DR11 had antibodies to CMV gH than did those without HLA-DR10 and DR11. HLA-DR10 and DR11 may be associated with fewer/non-responders for strain-specific neutralizing antibodies.     

The rate of human herpes virus type 6 and cytomegalovirus among HIV-infected and healthy women from Chelyabinsk and Moscow

Human herpes virus type 6 (H HV-6) and cytomegalovirus (CMV) seroprevalence among HIV-infected and healthy women of childbearing age from Chelyabinsk and Moscow was measured with ELI - SA test-system for detection of antibodies to respective viruses. Seropositivity to HHV-6 was detected in 93.5% of HIV-infected women, that is 1.2 times more often than in healthy woman of child-bearing age. Seropositivity to CMV was detected in 87% of HIV-infected women, that is 1.05 times more often than in healthy woman of childbearing age. Low titers to HHV-6 and CMV were detected in seropositive healthywomen more frequently (in 44.1 and 48.4% respectively) whereas intermediate and high titers prevailed in HIV-infected women (in 73.9% for HHV-6 and 31.4% for CMV).     

Cytomegalovirus IgM Seroprevalence among Women of Reproductive Age in the United States

Cytomegalovirus (CMV) IgM indicates recent active CMV infection. CMV IgM seroprevalence is a useful marker for prevalence of transmission. Using data from the National Health and Nutrition Examination Survey (NHANES) III 1988–1994, we present estimates of CMV IgM prevalence by race/ethnicity, provide a comparison of IgM seroprevalence among all women and among CMV IgG positive women, and explore factors possibly associated with IgM seroprevalence, including socioeconomic status and exposure to young children. There was no difference in IgM seroprevalence by race/ethnicity among all women (3.1%, 2.2%, and 1.6% for non-Hispanic white, non-Hispanic black and Mexican American, respectively; P = 0.11). CMV IgM seroprevalence decreased significantly with increasing age in non-Hispanic black women (P<0.001 for trend) and marginally among Mexican American women (P = 0.07), while no apparent trend with age was seen in non-Hispanic white women (P = 0.99). Among 4001 IgG+ women, 118 were IgM+, resulting in 4.9% IgM seroprevalence. In IgG+ women, IgM seroprevalence varied significantly by age (5.3%, 7.3%, and 3.7% for women of 12–19, 20–29, and 30–49 years; P = 0.04) and race/ethnicity (6.1%, 2.7%, and 2.0% for non-Hispanic white, non-Hispanic black, and Mexican American; P<0.001). The factors reported associated with IgG seroprevalence were not associated with IgM seroprevalence. The patterns of CMV IgM seroprevalence by age, race/ethnicity, and IgG serostatus may help understanding the epidemiology of congenital CMV infection as a consequence of vertical transmission and are useful for identifying target populations for intervention to reduce CMV transmission.     

T and B cell responses to cytomegalovirus antigens in healthy blood donors and bone marrow transplant recipients

Abstract We measured the production of interferon-gamma (IFN-γ) from single T cells and the T cell proliferative response to different cytomegalovirus (CMV) antigens in healthy blood donors and bone marrow transplant recipients. The antigens consisted of a CMV nuclear antigen (CMV na) containing the pp65-kDa matrix protein and the immediate early antigens but lacking CMV glycoproteins, and an antigen comprising native CMV glycoproteins (CMV gp). We also measured the IgG antibodies to CMV na and CMV gp. The T cells reacted to CMV na in CMV seropositive blood donors both with the production of IFN-γ and with proliferation, while bone marrow transplant recipients had a deficient T cell response. After stimulation with CMV gp, no T cell response could be observed in CMV seropositive subjects. IgG antibodies to CMV na coexisted in plasma with similar levels of antibodies to CMV gp.     

Co-electroporation of rice protoplasts with RNAs of cucumber mosaic and tobacco mosaic viruses

Cucumber mosaic virus (CMV) RNA was used to study electroporation conditions suitable for protoplasts from rice suspension cultures. Rice protoplasts required a stronger and shorter electric pulse than tobacco protoplasts for introduction of viral RNA. Under optimized conditions, CMV infection was established in 65 % of electroporated protoplasts. In contrast, electroporation with tobacco mosaic virus (TMV) RNA did not result in infection of rice protoplasts. However, when TMV RNA was electroporated into rice protoplasts together with CMV RNA, TMV production was demonstrated in 15 % of protoplasts. Differential staining with fluorescent antibodies against the two viruses showed that the protoplasts producing TMV were without exception also infected by CMV. The results show that CMV replicates in rice protoplasts by itself, whereas TMV does so only with the aid of CMV.Abbreviations CMV cucumber mosaiv virus - PBS phosphate buffered saline - TMV tobacco mosaic virus.     

Studies on the presence of antibodies to EB virus and other herpesviruses in normal children and in infectious mononucleosis.

Children free from infectious disease have been examined by indirect immunofluorescence for the presence of antibodies to the intracellular capsid antigen of Epstein-Barr virus (EBV). In the first year of life 46%, between 2 and 6 years of age 66%, and between 7 and 14 years 91%, of the children proved positive. The corresponding percentages for the presence of antibodies to cytomegalovirus (CMV) and herpes simplex virus were about 50%, irrespective of the children's age. Serum samples from 69 patients suffering from infectious mononucleosis (IM) were tested for anti-EBV antibodies. Of the 29 Paul-Bunnell-positive patients 22 had antibodies, 11 of them in high titres (greater than 1 : 80). Of the 40 Paul-Bunnell-negative cases only 21 had antibodies, 8 in high titres. Of the Paul-Bunnell-negative cases, 73% were found to have anti-CMV antibodies, 32% in high titre. The respective percentages for the Paul-Bunnell-positive cases were 42% and 10%.