Postinfection and postvaccination antirubella immunity

Out of 1670 pregnant women contacts examined for antibodies to rubella virus, recent rubella infection was confirmed in 5.1% of subjects, 2.2% of which were symptomatic and 2.9% asymptomatic; 89.9% of tested women were seropositive. These results closely resembled those, obtained in serological surveys, carried out in Poland several times in the years 1969-82. Rubella antibodies were found in over 90% of women in childbearing age, with HI antibody geometric mean titre (GMT) 58.7-84.3 (depending on the epidemiologic situation). Especially high GMT--of 122--was detected in women of the 30-34 year age group after the rubella epidemic in 1980/81, which showed the highest rubella morbidity of 5-9 year-old children. Reactogenicity and immune response in 10-13 year-old vaccinated girls were investigated. Seroconversion among seronegatives, among subjects with low antibody level (less than 1:20) and among seropositives with antibody titres 40 or higher before vaccination occurred in 99.8%, 60% and 12.8% of cases, respectively. HI antibody GMT after immunization was 132.6-135, similar to GMT induced by naturally acquired infection in the women of 30-34 year age group after the rubella epidemic. The difference of GMT in vaccinated girls with joint symptoms (152) and without postvaccinal reactions (124) was statistically significant. Practical conclusions for rubella diagnostic and vaccination programmes based on the analogies and differences of antirubella immunity in infected pregnant women, in naturally infected children and adults, and in vaccinated subjects are discussed.     

Rubella prevalence and its transmission in children

Rubella is a major cause of birth defects among the TORCH group of agents causing congenital anomalies. Almost all the symptomatic infected infants have long-term neurological sequelae & many asymptomatic infants also develop deafness or psychomotor retardation later in life. In India need for rubella prevention & control is being recognized. Before formulating any kind of rubella vaccination policies, data on the burden of disease is important. Hence the prevalence of rubella in children and their transmission was evaluated. Paired sera of 146 babies with suspected intra uterine infection and their mothers from lower socioeconomic strata was tested for IgM antibodies by commercially available Enzyme immunoassay (EIA) kits. Congenital Rubella Syndrome (CRS) was confirmed in babies presenting with rubella compatible defects with positive IgM antibodies against rubella. It was seen that out of 146-paired samples evaluated, 15-paired samples (10.27%) were positive for IgM antibodies. The transmission rate of rubella virus from mother to child when the mother was infected was around 55.55% according to this study. CRS prevalence of 10.27% among symptomatic infants is significant as a large majority of rubella infection remains undetected and hence the actual burden of the disease may be higher. Since the disease is preventable by an effective vaccination, strategies for rubella immunization should be developed and enhanced.     

Kinetics of rubella-specific IgM antibody response in postnatal rubella infection

The serological diagnosis of primary postnatal rubella infection is based on detection of rubella-virus-specific IgM antibody or a four-fold rise in rubella-specific IgG antibody. Although there are several different methods of enzyme immunoassays that are commercially available, the cost benefit evaluation makes them impractical for use in developing countries. For this reason, we have standardized the measurement of rubella IgM antibody by HAI following serum fractionation by ion-exchange chromatography. The sera samples obtained from pregnant women infected with rubella virus at different times during gestation were fractionated and tested by HAI. Seven out of nine sera collected within the first two days after onset of rash showed detectable levels of rubella IgM antibody. All 57 sera collected between 3 and 30 days after the onset of rash contained rubella IgM antibody. After 30 days, only 1 of 5, or 20%, of sera contained IgM antibody. The HAI testing method was rapid and specific and the cost was not prohibitive. HAI-IgM testing could be used to diagnose primary rubella infections in developing countries where expensive EIAs are unaffordable.     

Rubella vaccine: new horizon in prevention of congenital rubella syndrome in the India

Rubella is a contagious viral disease, which mainly affects the fetus, if the mother is infected in the 1st trimester of her pregnancy. All adolescent girls (aged 11 to 19 y) and women of childbearing age are at risk of developing rubella. This disease is mild and self-limiting, and incubation period is 2-3 weeks. Humans are the only hosts for rubella. Rubella infection during pregnancy may lead to abortions, stillbirth or congenital deformities (birth defects). Moreover it is surprising to know that over 200,000 babies are born with birth defects because of Rubella infection during pregnancy in the Indian sub-continent. The risk of fetal infection is highest in first trimester; the infection rate declines between 12-28 weeks, suggesting that the placenta may prevent transfer of virus but not completely. The incidence of defects is inversely related to the time of maternal infection. Rubella outbreaks have been reported from many countries in South East Asian region with congenital rubella syndrome (CRS) due to maternal rubella being on the increase in many countries. In India, although the endemicity of rubella is established, the majority of cases remain undiagnosed, being subclinical or clinically mild. Consequently, in spite of evidence of CRS in all States of India, no distinct policy has been envisaged for assessing the burden of rubella, and no control measures against this silent crippling disease are in place. The European Regional Committee of the World Health Organization has adopted the goals of "Elimination of CRS" in the Health for All programs. There is no treatment for rubella. Vaccination is the only way to prevent all these complications.     

Persistence and Titer Changes of Rubella Virus Antibodies in Primiparous Women Who Had Been Vaccinated with Strain RA 27/3 in Junior High School

Taiwan''s rubella vaccination program was launched in 1986; each schoolgirl in the third grade of junior high school received one dose of rubella (RA 27/3) vaccine. We reviewed the results of 14,090 prenatal rubella tests for primiparas from three areas of Taiwan during 2002 to 2008 to investigate seronegativity rates and titer changes. In all primiparous women, the average rubella virus seronegativity rate was 6.5% (95% confidence interval [95% CI], 6.1 to 6.9%), and the average rubella virus antibody titer was 65.9 IU/ml (95% CI, 64.7 to 67.1 IU/ml). There were 1,220 women (8.7%) with weakly positive antibody titers (10 to 20 IU/ml). The rubella virus seronegativity rates, which ranged from 5.4 to 9.7%, did not exhibit a linear trend from 9 to 22 years after vaccination (P = 0.201); in contrast, a significant trend appeared in the average rubella virus IgG titer (P = 0.003), dropping from 69.9 IU/ml in the 9th year after vaccination to 54.8 IU/ml in the 22nd year. The mean annual antibody decay rate was −0.77 IU/ml. This study reveals that the level of rubella virus antibodies declined slowly in women of childbearing age who were vaccinated with RA 27/3 at junior high school age. The number of women who were seronegative or had weakly positive antibody titers was still high (15.2%). Therefore, in countries that implement a single-dose regimen in children or teenagers, it should remain an important policy to encourage voluntary immunization in seronegative women and to immunize all postpartum women who are susceptible to rubella virus infection before they leave the hospital.     

High-Throughput Assay Optimization and Statistical Interpolation of Rubella-Specific Neutralizing Antibody Titers

Rubella remains a social and economic burden due to the high incidence of congenital rubella syndrome (CRS) in some countries. For this reason, an accurate and efficient high-throughput measure of antibody response to vaccination is an important tool. In order to measure rubella-specific neutralizing antibodies in a large cohort of vaccinated individuals, a high-throughput immunocolorimetric system was developed. Statistical interpolation models were applied to the resulting titers to refine quantitative estimates of neutralizing antibody titers relative to the assayed neutralizing antibody dilutions. This assay, including the statistical methods developed, can be used to assess the neutralizing humoral immune response to rubella virus and may be adaptable for assessing the response to other viral vaccines and infectious agents.     

Evaluation of a rapid passive hemagglutination assay for anti-rubella antibody: comparison to hemagglutination inhibition and a vaccine challenge study

A rapid passive hemagglutination assay (Rubaquick) was developed that detects antibody to rubella virus in serum specimens. The test result is read visually after an incubation period of 15-30 minutes. When compared with a hemagglutination inhibition assay, the Rubaquick assay results obtained from 1,470 sera were greater than 99% specific, sensitive, and accurate. Studies of 179 paired serum specimens obtained before and 27 days after rubella vaccination showed that if antibody was detectable by the Rubaquick assay in the prevaccination specimens, the vaccine induced a secondary response consisting of increasing IgG antibody reactivity in the absence of a positive IgM response. In contrast to the positive prevaccination specimens, a negative prevaccination result was associated with IgM antibody in 98 of the 133 postvaccination specimens. Seroconversion was noted in all cases in which the prevaccination specimen was negative by the Rubaquick assay.     

The present situation and limitation of antibody assays for diagnosis of rubella virus infection in pregnant women]

Rubella virus infection during early stages of pregnancy often results in a number of developmental disorders referred to as congenital rubella syndrome(CRS). Both clinical and laboratory diagnosis of suspect cases of CRS can be made with relative ease, particularly when expectant mothers show the typical rubella-specific rash. Serological diagnosis of CRS is accomplished using hemagglutination inhibition (HI) and enzyme-linked immunosorbent(IgM-EIA) assays. Antibody titers as determined by these assays are generally very high following acute apparent rubella infections, thus making serological diagnosis relatively easy in most cases. However, the detection of possible CRS cases can be hampered by clinically inapparent rubella infections during early pregnancy. As much as 30 percent of all acute rubella cases are inapparent infections, and there is the very real potential for such inapparent infections to occur during pregnancy, to result in fetal infections, and consequently to cause CRS. Detection of CRS becomes extremely difficult in such settings. Complicating CRS detection even more are rare rubella re-infections that might occur in early pregnancy, and unknown risk of fetal infection and CRS. In re-infection cases, HI antibody titer becomes elevated due to a secondary immune response, and IgM antibody is produced in a significant number of cases. To determine directly the fetal infection, virus genome detection was developed and applied clinically for the past decade. Using a combination of serological and genomic detection methods, the results of the investigation suggest that when rubella infection during early pregnancy occurs 1) there is a significant risk of fetal infection that results from acute apparent rubella infection, 2) there is a measurable risk of fetal infection resulting from inapparent infections as defined by HI antibody titers > or = 256 and with and IgM-EIA index > or = 7, and 3) high HI antibody titers with low IgM-EIA indices or no detectable IgM antibody in cases of inapparent rubella infections may represent rubella re-infections and result in a low risk of fetal infections.     

Cell-mediated immune response following natural rubella and rubella vaccination

Cell-mediated immunity (CMI), determined by estimating the production of MIF from sensitized leucocytes, was followed for 12 months after natural rubella infection (twenty-two subjects) and after vaccination with the Cendehill strain of attenuated rubella virus vaccine (forty subjects). Results were compared and correlated with the development of rubella haemagglutination inhibition antibody. The cell-mediated immune response commenced 1 week before the humoral immune response, but both responses reached a maximum simultaneously whether induced by natural infection or by vaccination. However, the CMI following natural rubella infection was of greater magnitude and duration than that stimulated by the vaccine virus. Twelve months after the initial stimulus, CMI could not be detected in any of the vaccinees, but was still present in some individuals after natural infection. This ability of the Cendehill vaccine virus to stimulate only shortlived, detectable CMI may be an important factor in the high reinfection rate observed following rubella vaccination.     

Rubella virus replication and links to teratogenicity

Rubella virus (RV) is the causative agent of the disease known more popularly as German measles. Rubella is predominantly a childhood disease and is endemic throughout the world. Natural infections of rubella occur only in humans and are generally mild. Complications of rubella infection, most commonly polyarthralgia in adult women, do exist; occasionally more serious sequelae occur. However, the primary public health concern of RV infection is its teratogenicity. RV infection of women during the first trimester of pregnancy can induce a spectrum of congenital defects in the newborn, known as congenital rubella syndrome (CRS). The development of vaccines and implementation of vaccination strategies have substantially reduced the incidence of disease and in turn of CRS in developed countries. The pathway whereby RV infection leads to teratogenesis has not been elucidated, but the cytopathology in infected fetal tissues suggests necrosis and/or apoptosis as well as inhibition of cell division of critical precursor cells involved in organogenesis. In cell culture, a number of unusual features of RV replication have been observed, including mitochondrial abnormalities, and disruption of the cytoskeleton; these manifestations are most probably linked and play some role in RV teratogenesis. Further understanding of the mechanism of RV teratogenesis will be brought about by the investigation of RV replication and virus-host interactions.     

Agreement of rubella IgG antibody measured in serum and dried blood spots using two commercial enzyme-linked immunosorbent assays

Cases of congenital rubella are now rare in the United Kingdom. However, in certain areas such as London, where a significant proportion of pregnant women has been born abroad and uptake of trivalent measles-mumps-rubella (MMR) vaccination is low, the risk of a rubella outbreak remains. Prior to carrying out a seroprevalence study using rubella IgG antibody in newborn dried blood spots as an indirect marker of maternal immunity, rubella IgG antibody concentrations in serum and dried blood spot samples were investigated. Anonymous paired serum-dried blood spot samples left over from occupational health screening were tested for rubella IgG antibody by two commercially available enzyme-linked immunosorbent assays (ELISAs) (Dade Behring, Marburg, Germany, and Diesse, Siena, Italy). Agreement between serum samples and dried blood spot samples was high for both assays. There were no significant differences in antibody concentrations in paired samples, as 67 of 73 samples tested with the Diesse ELISA (91.8%), and 76 out of 79 samples tested with the Dade Behring ELISA (96.2%) were within two standard deviations of the mean difference. Commercial ELISAs are an appropriate test for seroprevalence surveys based on rubella IgG in dried blood spot samples.     

The role of rubella-immunoblot and rubella-peptide-EIA for the diagnosis of the congenital rubella syndrome during the prenatal and newborn periods

Rubella infection during the first trimester of pregnancy can cause the congenital rubella syndrome (CRS). Patients with CRS were shown to have a decreased humoral and cellular immunity. It is not known whether asymptomatic newborns who had experienced intrauterine infection with rubella virus (RV) differ in their antibody response from newborns with CRS. In this study we compared both groups for a difference which might be a useful diagnostic criterion for CRS during the prenatal and newborn periods. We used the nonreducing Rubella-Immunoblot and the Rubella-IgG-Peptide-Enzyme Immunoassay (EIA) to determine the antibodies directed to rubella proteins E1, E2 and C. The results showed that only newborns with CRS who had experienced RV infection during the first 12 weeks of gestation showed significantly reduced levels of antibodies directed to both the linear RV E1 epitope (SP 15) and the topographic RV E2 epitope. Asymptomatic newborns infected mostly later than week 10 of gestation showed normal levels of antibodies. These data suggest that the lack of antibody response in CRS is linked to the immaturity of the fetal immune system during the first trimester of gestation. Rubella-IgG-Peptide-EIA and Rubella-Immunoblot should be used additionally for CRS diagnosis in the prenatal/newborn periods. These results may have an impact on the early treatment of late-onset symptoms of CRS patients. J. Med. Virol. 51:280–283, 1997. © 1997 Wiley-Liss, Inc.     

Venezuelan equine encephalitis-specific immunoglobulin responses: live attenuated TC-83 versus inactivated C-84 vaccine.

Venezuelan equine encephalitis (VEE)-specific immunoglobulin responses to the two vaccines, TC-83 (a live attenuated vaccine) and C-84 (a formalin inactivated vaccine derived from the TC-83 strain of virus) were evaluated using an antigen and isotype-specific enzyme-linked immunoadsorbent assay (ELISA). The VEE-specific ELISA for IgG, IgG subclasses, IgA and IgM were developed and standardized using sera from vaccine-exposed and unexposed human subjects. Paired human sera (before and 28 days after immunization) were tested from laboratory workers vaccinated with either TC-83 (Group A: 20 paired sera from subjects receiving a single TC-83 vaccine and with no prior history of vaccination) or C-84 in varying schedules (Group B: 19 paired sera from subjects who had a distant vaccination history to TC-83 but no evidence of neutralizing antibody; Group C: 19 paired sera from subjects receiving their first C-84 vaccination and no prior documented history of vaccination; Group D: 15 paired sera from subjects receiving a C-84 booster vaccination with prior history of C-84 but no TC-83 exposure). Sera were all tested for viral neutralization in vitro using a Vero cell monolayer for culturing virus and establishing 80% plaque reduction for each serum tested. All pre-sera tested demonstrated no plaque reduction neutralization at a level of 80% for a dilution of 1:10. ELISA antibody titers for all pre-sera with no prior VEE exposure through vaccination or possible environmental factors were negative at a titer of 1:160 for IgM, 1:80 for IgG, IgA, and G subclasses.(ABSTRACT TRUNCATED AT 250 WORDS)     

Evaluation of a commercial rubella IgM assay for use on oral fluid samples for diagnosis and surveillance of congenital rubella syndrome and postnatal rubella.

BACKGROUND: Clinical diagnosis (surveillance) of rubella is unreliable and laboratory confirmation is essential. Detection of virus specific IgM in serum is the most commonly used method. However, the use of serum necessitates the drawing of blood, either through venipuncture or finger/heel prick, which can be difficult in young babies. Oral fluid samples have proved useful as an alternative, less invasive sample for virus specific IgM detection however until recently no commercial rubella IgM tests were available, restricting the usefulness of this approach. OBJECTIVES: To evaluate the performance of the Microimmune Rubella IgM capture EIA using oral fluid samples from outbreaks as well as in cases of suspected congenital rubella syndrome (CRS). STUDY DESIGN: Paired serum and oral fluids were collected from cases during a rubella outbreak in three provinces in Turkey. Matched serum and oral fluid samples were collected from children with suspected CRS in an active surveillance programme at the Aravind Eye Hospital in South India. Serum samples were collected as part of the measles surveillance programme in Ethiopia. RESULTS: On serum samples the sensitivity and specificity of the Microimmune Rubella IgM capture EIA compared to Behring Enzygnost rubella IgM test was 96.9% (62/64; 95% CI 94.2-100%) and 100% (53/53; 95% CI 93.2-100%). On oral fluids compared to matched Behring results on serum the sensitivity was 95.5% (42/44; 95% CI 84.5-99.4%). The sensitivity and specificity of Microimmune Rubella IgM capture EIA on oral fluids from suspected CRS cases compared to serum results using Behring Enzygnost IgM assay was 100% (95% CI 84.5-100%) and 100% (95% CI 95.8-100.0%) respectively. CONCLUSION: Microimmune Rubella IgM capture EIA has adequate performance for diagnosis and surveillance of rubella in outbreak using either serum or oral fluid specimens.     

Transcriptional signatures associated with rubella virus-specific humoral immunity after a third dose of MMR vaccine in women of childbearing age

Multiple factors linked to host genetics/inherent biology play a role in interindividual variability in immune response outcomes after rubella vaccination. In order to identify these factors, we conducted a study of rubella-specific humoral immunity before (Baseline) and after (Day 28) a third dose of MMR-II vaccine in a cohort of 109 women of childbearing age. We performed mRNA-Seq profiling of PBMCs after rubella virus in vitro stimulation to delineate genes associated with post-vaccination rubella humoral immunity and to define genes mediating the association between prior immune response status (high or low antibody) and subsequent immune response outcome. Our study identified novel genes that mediated the association between prior immune response and neutralizing antibody titer after a third MMR vaccine dose. These genes included the following: CDC34; CSNK1D; APOBEC3F; RAD18; AAAS; SLC37A1; FAS; and JAK2. The encoded proteins are involved in innate antiviral response, IFN/cytokine signaling, B cell repertoire generation, the clonal selection of B lymphocytes in germinal centers, and somatic hypermutation/antibody affinity maturation to promote optimal antigen-specific B cell immune function. These data advance our understanding of how subjects’ prior immune status and/or genetic propensity to respond to rubella/MMR vaccination ultimately affects innate immunity and humoral immune outcomes after vaccination.     

Current seroepidemiology of rubella virus infection among female residents in Taiwan

To examine the epidemiological characteristics of rubella virus infection on Taiwan Island and in Orchid and Pescadore islets, a community-based survey was carried out in 19 townships and metropolitan precincts randomly selected through stratified sampling. Serum specimens of 4,770 healthy girls and women randomly sampled from study areas were screened for the rubella antibody by passive latex agglutination testing and confirmed by enzyme-linked immunosorbent assay. A total of 2,934 subjects were antibody positive, giving a prevalence of 61.5%. The mean ± standard deviation of rubella IgG index was 2.51 ± 0.37. The seropositive rate increased with age in all residential areas and ethnic groups. The age pattern indicated that the regular 10-year cycle of rubella epidemic was no longer existent after 1978. A higher prevalence was observed in the eastern island and Orchid Islets compared with other areas. The highest seropositive rate was found in Orchid Islets. The overall seronegative rate was 62.9% for females <20 years of age and 36.9% for women between the ages of 20 and 29 years. With such a high susceptible density among girls and women of child-bearing ages as well as the endemicity of rubella virus infection in Taiwan, mass vaccination against rubella should be enforced in order to prevent possible future outbreaks of the congenital rubella syndrome.     

Cellular and humoral immune responses to measles in immune adults re-immunized with measles vaccine

The objective of this study was to characterize the kinetics of the cellular and humoral immune responses elicited by measles vaccine given to previously immune adults. The cellular and humoral immune responses to measles were measured in seven healthy adults, before vaccination and at 1, 2, 3, and 4 weeks and 3 months after vaccination, using measles-specific T-cell proliferation and plaque reduction neutralization assays. All study subjects had detectable measles antibodies, but only six (85%) showed protective titers, defined as >1:120, before immunization. However measles-specific T-cell proliferation was not detectable before vaccination in any of the subjects. The six subjects with protective titers showed a positive stimulation index (SI) of >3.0 within the first 4 weeks after vaccination, an SI of 5 at the 4th week, and an SI of 3 at 3 months after vaccination. The subject with a low antibody titer (1:99) before vaccination developed a high SI at 3 months after vaccination. This subject was the only participant whose neutralizing antibody titers increased more than 4-fold by 3 months after vaccination. No significant increases in geometric mean titers were detected in the other six subjects during the follow-up period. These data suggest that high measles antibody titers interfere with the humoral response in subjects who receive a booster immunization, whereas the cellular response is boosted at least transiently, after revaccination.     

Antibody response to wild rubella virus structural proteins following immunization with RA 27/3 live attenuated vaccine

Summary Antibody response to individual structural proteins (E 1, E 2, and C) of the M-33 wild rubella virus strain was assayed by an immunoblot technique in 12 girls, following immunization with RA 27/3 live attenuated rubella vaccine. Of the 12 immunized subjects, before vaccination 9 had no demonstrable rubella specific antibodies while the remaining 3 had a low level of rubella specific antibodies, reacting only with the E 1 protein. At one month after vaccination all the immunized subjects presented anti-E 1, anti-E 2, and anti-C specific antibodies. However, at 1–2 weeks after vaccination the 9 girls who were seronegative before immunization still had no detectable antibodies to any of the rubella virus structural proteins, while the 3 subjects whose preimmunization sera had reacted with the E 1 protein presented an accelerated immune response, showing anti-E 2 and anti-C specific antibodies and a more intensely marked anti-E 1 specific band.     

Antibody response to rubella virion (V) and soluble (S) antigens in rubella infection and following vaccination with live attenuated rubella virus

Summary The antibody response to rubella virion antigen and rubella S antigen was studied in natural rubella infection and after vaccination with live attenuated rubella virus by the complement fixation (CF), platelet aggregation (PA), and hemagglutination inhibition (HI) techniques.In natural rubella infection CP and HI antibodies to rubella virion appeared early and rapidly reached high titers. The CP and PA antibody responses to rubella S antigen were more delayed and great individual variation was seen. Generally CF-S titers were several times lower than CF-V titers. The CF antibody response to rubella S antigen is thus different from the immune response to nucleoprotein S antigens of paramyxoviruses, supporting the concept that rubella S antigen is a subunit of virus envelope. Use of virus-free rubella S antigen preparations in routine CF test is recommended for detecting later rises of antibody titer.After rubella vaccination a 95% seroconversion rate was recorded in both HI and CF-V tests, but the titers were lower than after natural infection. The CF-S and PA antibody responses were weaker and measurable antibodies developed in only 50% and 25% of the cases respectively.Rubella-specific IgM antibodies could be detected in rubella infection by both sucrose gradient fractionation (followed by HI titration) and fluorescent antibody (FA) techniques. The former was a little more sensitive. In the seronegative vaccinees IgM antibodies became demonstrable in 50% of the cases between the 20th and 55th day after vaccination.