Virus-specific Immunoglobulin G Research Articles

Early-life exposure to perfluoroalkyl substances and serum antibody concentrations towards common childhood vaccines in 18-month-old children in the Odense Child Cohort

Exposure to per- and polyfluoroalkyl substances (PFAS) has been associated with reduced antibody response to childhood vaccinations. Previous studies have mostly focused on antibodies against diphtheria or tetanus, while fewer studies have assessed antibodies toward attenuated viruses, such as measles, mumps or rubella (MMR). Therefore, we set out to determine associations between prenatal and early postnatal PFAS exposure and vaccine-specific Immunoglobulin G (IgG) in the background-exposed Odense Child Cohort. Blood samples were drawn in pregnancy at gestation weeks 8–16 and from the offspring at age 18 months. In the maternal serum samples we quantified perfluorooctane sulfonic acid (PFOS), perfluorooctanoic acid (PFOA), perfluorohexane sulfonic acid (PFHxS), perfluorononanoic acid (PFNA) and perfluorodecanoic acid (PFDA). In the offspring serum samples we quantified the same five PFAS compounds and IgG towards diphtheria, tetanus and MMR. A total of 880 and 841 children were included in the analyses of diphtheria and tetanus or MMR, respectively. Multiple linear regression models were used for estimation of difference in virus-specific IgG per doubling of PFAS concentrations. Maternal PFAS concentrations were non-significantly inversely associated with most vaccine-specific antibody concentrations. Likewise, child PFAS concentrations were associated with non-significant reductions of antibodies towards tetanus and MMR. A significant reduction in the percent difference in mumps antibody concentration per doubling of child PFNA (−9.2% (95% confidence interval: −17.4;-0.2)), PFHxS (−8.3% (−15.0;-1.0) and PFOS (−7.9% (−14.8;-0.4) was found. These findings are of public health concern, as inadequate response towards childhood vaccines may represent a more general immune dysfunction.

Effectiveness of immunization activities on measles and rubella immunity among individuals in East Sepik, Papua New Guinea: A cross-sectional study

ObjectivesThis study aimed to assess measles and rubella immunity by measuring virus-specific immunoglobulin G (IgG) prevalence among individuals and evaluate the effectiveness of recent supplementary immunization activities (SIAs) by comparing the antibody positivity rates of the SIA target age groups in 2015 with those in 2019 as measles and rubella are endemic in Papua New Guinea. MethodsA cross-sectional study. The measles- and rubella-specific IgG levels of patients aged ≥1 year at two clinics in East Sepik province, Papua New Guinea were assessed with commercially available virus-specific IgG EIA kits. ResultsIn total, 297 people participated in the study and 278 samples with sufficient volume, relevant information, and age inclusion criteria were analyzed. The overall IgG prevalence rates were 62.6% for measles and 82.0% for rubella. The age groups targeted in the 2019 SIAs had a higher IgG prevalence than those targeted in the 2015 SIAs for both the infectious diseases. Moreover, the IgG prevalence for rubella was higher than measles in these groups. ConclusionsThe anti-measles and anti-rubella IgG prevalence in the target groups were lower than those required for herd immunity. The immunization program should be emphasized to eliminate measles and rubella. Further population-based studies are warranted.

Kinetics and persistence of anti-SARS-CoV-2 neutralisation and antibodies after BNT162b2 vaccination in a Swiss cohort.

IntroductionSince the emergence of severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2), substantial effort has been made to gain knowledge about the immunity elicited by infection or vaccination.MethodsWe studied the kinetics of antibodies and virus neutralisation induced by vaccination with BNT162b2 in a Swiss cohort of SARS‐CoV‐2 naïve (n = 40) and convalescent (n = 9) persons. Blood sera were analysed in a live virus neutralisation assay and specific IgG and IgA levels were measured by enzyme‐linked immunoassay and analysed by descriptive statistics.ResultsVirus neutralisation was detected in all individuals 2–4 weeks after the second vaccine. Both neutralisation and antibodies remained positive for >4 months. Neutralisation and antibodies showed positive correlation, but immunoglobulin G (IgG) and immunoglobulin A (IgA) seroconversion took place 2–4 weeks faster than neutralisation. Spike‐protein specific IgG levels rose significantly faster and were more stable over time than virus neutralisation titres or IgA responses. For naïve but not convalescent persons, a clear boosting effect was observed. Convalescent individuals showed faster, more robust and longer‐lasting immune responses after vaccination compared to noninfected persons. No threshold could be determined for spike protein‐specific IgG or IgA that would confer protection in the neutralisation assay, implicating the need for a better correlate of protection then antibody titres alone.ConclusionsThis study clearly shows the complex translation of antibody data and virus neutralisation, while supporting the evidence of a single dose being sufficient for effective antibody response in convalescent individuals.

Anti‐SARS‐CoV‐2 IgG levels in relation to disease severity of COVID‐19

The durability of infection‐induced severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) immunity has crucial implications for reinfection and vaccine effectiveness. However, the relationship between coronavirus disease 2019 (COVID‐19) severity and long‐term anti‐SARS‐CoV‐2 immunoglobulin G (IgG) antibody level is poorly understood. Here, we measured the longevity of SARS‐CoV‐2‐specific IgG antibodies in survivors who had recovered from COVID‐19 1 year previously. In a cohort of 473 survivors with varying disease severity (asymptomatic, mild, moderate, or severe), we observed a positive correlation between virus‐specific IgG antibody titers and COVID‐19 severity. In particular, the highest virus‐specific IgG antibody titers were observed in patients with severe COVID‐19. By contrast, 74.4% of recovered asymptomatic carriers had negative anti‐SARS‐CoV‐2 IgG test results, while many others had very low virus‐specific IgG antibody titers. Our results demonstrate that SARS‐CoV‐2‐specific IgG persistence and titer depend on COVID‐19 severity.

M408 AN UNUSUAL MISSED DIAGNOSIS OF MULTIFACETED CLINICAL CASE OF CORONAVIRUS DISEASE 2019 MIMICKING UNCONTROLLED ASTHMA

A patient with progressive shortness of breath, dry cough, tightness of the chest, mimic uncontrolled asthma. A diagnosis of coronavirus disease 2019 (COVID-19) infection might be delayed if a chest CT scan is not done (after abnormality of chest X-ray), despite a negative nasopharyngeal swab test reverse transcription, polymerase chain reaction (RT-PCR) assay. A 68-year old female presented to us with progressive shortness of breath worsening over 12 days, a dry cough, and tightness of the chest. Her primary care diagnosed her with severe persistent asthma and prescribed an inhaled bronchodilator, corticosteroid, anticholinergic and montelukast with a course of oral corticosteroid. The patient came to our clinic due to the worsening of symptoms despite the step-up therapy for asthma. Physical examination revealed decreased airflow and bilateral rales in both lungs, blood pressure: 137/78, temperature 38.9 C, heart rate 109 per minute, oxygen saturation 90 %. The chest CT Scan showed multiple ground-glass opacities and the serological antibodies tests against SARS-COV-2 were found positive for immunoglobulin M (IgM) and immunoglobulin G (IgG), confirming the diagnosis of COVID-19. Various medical conditions may mimic uncontrolled asthma. Patients unresponsive to asthma treatment should be evaluated for the presence of an asthma mimic, in this case COVID-19 infection. A chest CT Scan should be performed on the patient with serological virus specific IgG and IgM antibodies against SARS-COV-2 to diagnosis COVID-19 despite the negativity of the RT-PCR assay. This case highlights the complexity and the multifaceted presentation of coronavirus disease 2019 (COVID-19).

Measles and rubella serosurvey identifies rubella immunity gap in young adults of childbearing age in Zambia: The added value of nesting a serological survey within a post-campaign coverage evaluation survey

BackgroundSerological surveys can potentially complement vaccine coverage surveys, such as post-vaccination campaign coverage evaluation surveys (PCES), by providing direct information on population immunity within and outside the target age range of the mass vaccination campaign. We estimate age-specific population immunity to measles and rubella viruses in Southern Province, Zambia, and assess the value of adding serological data to vaccination coverage estimates by nesting a serological survey within a PCES. MethodsDried blood spots (DBS) from fingerprick blood were collected from all individuals ages nine months or older in households participating in the PCES and tested for measles and rubella virus-specific immunoglobulin G (IgG) by enzyme immunoassay (Siemens Enzygnost, Marburg, Germany). ResultsOverall seroprevalence was 95.5% (95% CI: 92.8, 97.2) for measles virus-specific IgG and 97.7% (95% CI: 96.0, 98.7) for rubella virus-specific IgG. Rubella seroprevalence was 98.4% (95% CI: 95.9, 99.4) among children eligible for the MR vaccination campaign, significantly higher than the reported measles-rubella (MR) vaccination campaign coverage of 89.8% (p = 0.003), and higher than the 91.3% rubella seroprevalence for adolescents and adults 16–30 years of age (p = 0.049). ConclusionSeroprevalence to measles and rubella viruses in children younger than 16 years of age was significantly higher than expected from vaccination coverage estimates, likely reflecting exposure to wild-type viruses and underreporting of vaccination. The serosurvey revealed rubella immunity gaps among women 16–30 years of age, precisely the age group in which protection from rubella is most important to prevent congenital rubella syndrome. Nesting serological surveys within existing surveys can leverage resources and infrastructure while providing complementary information important to immunization programs.

Adaptive Immune Responses in Humans During Nipah Virus Acute and Convalescent Phases of Infection.

Nipah virus (NiV) is 1 of 10 potential causes of imminent public health emergencies of international concern. We investigated the NiV outbreak that occurred in May 2018 in Kerala, India. Here we describe the longitudinal characteristics of cell-mediated and humoral immune responses to NiV infection during the acute and convalescent phases in 2 human survivors. Serial blood samples were obtained from the only 2 survivors of the NiV outbreak in Kerala. We used flow cytometry to determine the absolute T-lymphocyte and B-lymphocyte counts and the phenotypes of both T and B cells. We also detected and quantitated the humoral immune response to NiV by virus-specific immunoglobulin M (IgM) and immunoglobulin G (IgG) enzyme-linked immunosorbent assay. Absolute numbers of T lymphocytes remained within normal limits throughout the period of illness studied in both survivors. However, a marked elevation of activated CD8 T cells was observed in both cases. More than 30% of total CD8 T cells expressed Ki67, indicating active proliferation. Proliferating (Ki-67+) CD8 T cells expressed high levels of granzyme B and PD-1, consistent with the profile of acute effector cells. Total B-lymphocyte, activated B-cell, and plasmablast counts were also elevated in NiV survivors. These individuals developed detectable NiV-specific IgM and IgG antibodies within a week of disease onset. Clearance of NiV RNA from blood preceded the appearance of virus-specific IgG and coincided with the peak of activated CD8 T cells. We describe for the first time longitudinal kinetic data on the activation status of human B- and T-cell populations during acute NiV infection. While marked CD8 T-cell activation was observed with effector characteristics, activated CD4 T cells were less prominent.

A Historical Look at the First Reported Cases of Lassa Fever: IgG Antibodies 40 Years After Acute Infection

Lassa fever is an acute and sometimes severe viral hemorrhagic illness endemic in West Africa. One important question regarding Lassa fever is the duration of immunoglobulin G (IgG) antibody after infection. We were able to locate three persons who worked in Nigeria dating back to the 1940s, two of whom were integrally involved in the early outbreaks and investigations of Lassa fever in the late 1960s, including the person from whom Lassa virus was first isolated. Two persons had high titers of Lassa virus-specific IgG antibody over 40 years after infection, indicating the potential for long-term duration of these antibodies. One person was likely infected in 1952, 17 years before the first recognized outbreak. We briefly recount the fascinating stories of these three pioneers and their important contribution to our understanding of Lassa fever.

A Historical Look at the First Reported Cases of Lassa Fever: IgG Antibodies 40 Years After Acute Infection

Lassa fever is an acute and sometimes severe viral hemorrhagic illness endemic in West Africa. One important question regarding Lassa fever is the duration of immunoglobulin G (IgG) antibody after infection. We were able to locate three persons who worked in Nigeria dating back to the 1940s, two of whom were integrally involved in the early outbreaks and investigations of Lassa fever in the late 1960s, including the person from whom Lassa virus was first isolated. Two persons had high titers of Lassa virus-specific IgG antibody over 40 years after infection, indicating the potential for long-term duration of these antibodies. One person was likely infected in 1952, 17 years before the first recognized outbreak. We briefly recount the fascinating stories of these three pioneers and their important contribution to our understanding of Lassa fever.

NF-κB p50 Plays Distinct Roles in the Establishment and Control of Murine Gammaherpesvirus 68 Latency

NF-kappaB signaling is critical to the survival and transformation of cells infected by the human gammaherpesviruses Epstein-Barr virus and Kaposi's sarcoma-associated herpesvirus. Here we have examined how elimination of the NF-kappaB transcription factor p50 from mice affects the life cycle of murine gammaherpesvirus 68 (MHV68). Notably, mice lacking p50 in every cell type were unable to establish a sufficiently robust immune response to control MHV68 infection, leading to high levels of latently infected B cells detected in the spleen and persistent virus replication in the lungs. The latter correlated with very low levels of virus-specific immunoglobulin G (IgG) in the infected p50(-/-) mice at day 48 postinfection. Because the confounding impact of the loss of p50 on the host response to MHV68 infection prevented a direct analysis of the role of this NF-kappaB family member on MHV68 latency in B cells, we generated and infected mixed p50(+/+)/p50(-/-) bone marrow chimeric mice. We show that the chimeric mice were able to control acute virus replication and exhibited normal levels of virus-specific IgG at 3 months postinfection, indicating the induction of a normal host immune response to MHV68 infection. However, in p50(+/+)/p50(-/-) chimeric mice the p50(-/-) B cells exhibited a significant defect compared to p50(+/+) B cells in supporting MHV68 latency. In addition to identifying a role for p50 in the establishment of latency, we determined that the absence of p50 in a subset of the hematopoietic compartment led to persistent virus replication in the lungs of the chimeric mice, providing evidence that p50 is required for controlling virus reactivation. Taken together, these data demonstrate that p50 is required for immune control by the host and has distinct tissue-dependent roles in the regulation of murine gammaherpesvirus latency during chronic infection.

Development of a Rapid and Convenient Method for Determination of Rubella Virus-Specific Immunoglobulin G Avidity

We describe here a rapid and semiautomated method for the determination of rubella virus immunoglobulin G (IgG) avidity with the VIDAS instrument. A total of 153 serum samples from persons with naturally acquired rubella virus infections (n = 98), from vaccinated persons (n = 44), and from patients with autoantibodies (n = 11) were included in this study. The rubella virus-specific IgG avidity assay we developed for the VIDAS instrument was evaluated by comparison with an in-house method. Results obtained with the VIDAS instrument allow considering this method valuable to help confirm or exclude acute primary infection or recent vaccination.

Detection of Antibodies to Egg Drop Syndrome Virus in Chicken Serum Using a Field-Based Immunofiltration (Flow-Through) Test

A simple, user-friendly, and rapid method to detect the presence of antibodies to egg drop syndrome 76 (EDS) virus in chicken sera based on an immunofiltration (flow-through) test was developed. Purified EDS virus antigen was coated onto nitrocellulose membranes housed in a plastic module with layers of absorbent filter pads underneath. Following addition of serum to be tested and washing, monoclonal antibodies or polyclonal serum to chicken immunoglobulin G (IgG) was used as a bridge antibody to mediate binding between EDS virus-specific IgG and protein A gold conjugate. The appearance of a pink dot indicated the presence of antibodies to EDS virus in the sample tested. The results could be obtained within 5-10 min. The developed immunofiltration test could detect antibodies in the sera of experimentally vaccinated chickens from 2 wk postvaccination. With field sera samples, this test was positive in samples having hemagglutination inhibition titers of 8 and above. This test has the potential to be used as a field-based kit to assess seroconversion in EDS-vaccinated flocks.

Detection of virus-specific intrathecally synthesised immunoglobulin G with a fully automated enzyme immunoassay system

BackgroundThe determination of virus-specific immunoglobulin G (IgG) antibodies in cerebrospinal fluid (CSF) is useful for the diagnosis of virus associated diseases of the central nervous system (CNS) and for the detection of a polyspecific intrathecal immune response in patients with multiple sclerosis. Quantification of virus-specific IgG in the CSF is frequently performed by calculation of a virus-specific antibody index (AI). Determination of the AI is a demanding and labour-intensive technique and therefore automation is desirable. We evaluated the precision and the diagnostic value of a fully automated enzyme immunoassay for the detection of virus-specific IgG in serum and CSF using the analyser BEP2000 (Dade Behring).MethodsThe AI for measles, rubella, varicella-zoster, and herpes simplex virus IgG was determined from pairs of serum and CSF samples of patients with viral CNS infections, multiple sclerosis and of control patients. CSF and serum samples were tested simultaneously with reference to a standard curve. Starting dilutions were 1:6 and 1:36 for CSF and 1:1386 and 1:8316 for serum samples.ResultsThe interassay coefficient of variation was below 10% for all parameters tested. There was good agreement between AIs obtained with the BEP2000 and AIs derived from the semi-automated reference method.ConclusionDetermination of virus-specific IgG in serum-CSF-pairs for calculation of AI has been successfully automated on the BEP2000. Current limitations of the assay layout imposed by the analyser software should be solved in future versions to offer more convenience in comparison to manual or semi-automated methods.

Papillomavirus Capsid Mutation To Escape Dendritic Cell-Dependent Innate Immunity in Cervical Cancer

Infection with oncogenic human papillomaviruses (HPVs), typified by HPV type 16 (HPV16), is a necessary cause of cervical cancer. Prophylactic vaccination with HPV16 L1 virus-like particles (VLPs) provides immunity. HPV16 VLPs activate dendritic cells and a potent neutralizing immunoglobulin G (IgG) response, yet many cervical cancer patients fail to generate detectable VLP-specific IgG. Therefore, we examined the role of the innate recognition of HPV16 L1 in VLP-induced immune responses and its evasion during carcinogenesis. Nonconservative mutations within HPV16 L1 have been described in isolates from cervical cancer and its precursor, high-grade cervical intraepithelial neoplasia (CIN). We determined the effect of mutations in L1 upon in vitro self-assembly into VLPs and their influence upon the induction of innate and adaptive immune responses in mice. Several nonconservative mutations in HPV16 L1 isolated from high-grade CIN or cervical carcinoma prevent self-assembly of L1 VLPs. Intact VLPs, but not assembly-defective L1, activate dendritic cells to produce proinflammatory factors, such as alpha interferon, that play a critical role in inducing adaptive immunity. Indeed, effective induction of L1-specific IgG1 and IgG2a was dependent upon intact VLP structure. Dendritic cell activation and production of virus-specific neutralizing IgG by VLPs requires MyD88-dependent signaling, although the L1 structure that initiates MyD88-mediated signaling is distinct from the neutralizing epitopes. We conclude that innate recognition of the intact L1 VLP structure via MyD88 is critical in the induction of high-titer neutralizing IgG. Tumor progression is associated with genetic instability and L1 mutants. Selection for assembly-deficient L1 mutations suggests the evasion of MyD88-dependent immune control during cervical carcinogenesis.

Persisting Humoral Antiviral Immunity within the Japanese Population after the Discontinuation in 1976 of Routine Smallpox Vaccinations

Concerns have arisen recently about the possible use of smallpox for a bioterrorism attack. Routine smallpox vaccination was discontinued in Japan in 1976; however, it is uncertain exactly how long vaccination-induced immunity lasts. We sought to evaluate the seroprevalence and intensity of anti-smallpox immunity among representatives of the present Japanese population. The subjects included 876 individuals who were born between 1937 and 1982. Vaccinia virus-specific immunoglobulin G (IgG) levels were measured by enzyme-linked immunosorbent assay (ELISA), and 152 of 876 samples were also tested for the presence of neutralizing antibodies. Of the subjects who were born before 1962, between 1962 and 1968, and between 1969 and 1975, 98.6, 98.6, and 66.0%, respectively, still retained the vaccinia virus-specific IgG with ELISA values for optical density at 405 nm (OD(405)) of > or = 0.10. The corresponding figures for retained IgGs with OD405 values of > or = 0.30 were 91.0, 90.3, and 58.2%, respectively. Neutralizing antibodies were also maintained. The sera with OD(405) values of > or = 0.30 showed 89% sensitivity and a 93% positive predictive value for detection of neutralizing antibodies (> or = 4). Thus, approximately 80% of persons born before 1969 and 50% of those born between 1969 and 1975 were also found to have maintained neutralizing antibodies against smallpox. A considerable proportion of the previous vaccinated individuals still retain significant levels of antiviral immunity. This long-lasting immunity may provide some protective benefits in the case of reemergence of smallpox, and the disease may not spread as widely and fatally as generally expected.

Stability of total and rubella-specific IgG in oral fluid samples: the effect of time and temperature

Oral fluid (saliva) samples are increasingly used as a minimally invasive alternative to serum to detect antibody for viral diagnostics and population-based surveys of immunity to common infections. Postal collection of these samples is convenient, but it is uncertain how oral fluid antibody levels may deteriorate during transit. In this study multiple oral fluid samples, from a group of individuals, were collected and kept at two different temperatures (10 and 20 °C, simulating winter and summer conditions ) for periods of up to 7 days. They were then tested for total immunoglobulin G (IgG) and for rubella-specific IgG. Total IgG concentrations ranged from 2.0 to 48.6 mg/l but showed no significant decrease over the 7-day period. There was also no significant change in rubella-specific IgG measured using an antibody capture ELISA, but testing with an indirect rubella IgG ELISA revealed a significant decrease in absorbance values between day 1 and day 7 at 20 °C. The difference in the rubella assay results may reflect the different assay formats with the signal in the indirect procedure related to the concentration of rubella IgG in the oral fluid sample, rather than the proportion of virus-specific IgG measured with the antibody capture format. The study demonstrated that total and rubella specific IgG in oral fluid samples were stable for up to 7 days at differing temperatures and that an antibody capture assay format should be used for virus-specific testing to minimise inaccurate results due to low IgG concentrations in oral fluid samples.

Reconstitution of humoral immunity during pregnancy.

Recently, increasing attention has been paid to hormonal regulations of the immune system. In this study, cord sera and the corresponding maternal sera were obtained at delivery. Sera from pregnant women were obtained at early, middle, and late stages of pregnancy. These sera were tested for titer and avidity of measles or mumps virus-specific immunoglobulin G (IgG) by means of a single-dilution, urea-denaturation enzyme-linked immunosorbent assay method. A positive and significant correlation was found between the titer and avidity of the virus-specific IgG, both in the cord sera and in the maternal sera. This correlation was established already at the early stage of pregnancy. There were no such correlations found in nonpregnant individuals. This is the first observation in human subjects that the avidity and concentration of the virus-specific IgG had a positive and significant correlation. Pregnancy must have some significant effects on the regulation of humoral immunity.

Evaluation of Immunoglobulin M (IgM) and IgG Enzyme Immunoassays in Serologic Diagnosis of West Nile Virus Infection

A unique urban encephalitis epidemic in Romania signaled the emergence of neurological infection due to West Nile (WN) virus as a novel public health threat in Eastern Europe and provided an opportunity to evaluate patterns of immunoglobulin G (IgG) and IgM reactivity in IgM capture and IgG enzyme-linked immunosorbent assays (ELISAs). WN virus infection was diagnosed serologically in 236 of 290 patients from whom acute serum or cerebrospinal fluid (CSF) samples were available. In 37% of serum samples and in 25% of CSF samples collected in the first week of illness, anti-WN virus IgM antibody was detected in the absence of virus-specific IgG. The switch to an IgG antibody response occurred after 4 to 5 days of illness and earlier in CSF than in serum. A specific humoral immune response was detected in the CSF before the serum in some patients for whom paired CSF and serum samples from the same day were available. IgM antibody in convalescent serum samples persisted beyond 2 months after the onset of illness in more than 50% of patients. ELISA optical density values and antibody concentrations were well correlated for both IgM and IgG immunoassays. Anti-WN virus IgM antibody in acute-phase samples did not cross-react significantly with flaviviruses in other antigenic groups.

Detection of rubella virus-specific immunoglobulin G in saliva by an amplification-based enzyme-linked immunosorbent assay using monoclonal antibody to fluorescein isothiocyanate.

An immunoglobulin G (IgG)-capture enzyme-linked immunosorbent assay (ELISA) for rubella virus is described. The assay uses a fluorescein isothiocyanate (FITC)-anti-FITC amplification system. The detection limit of the ELISA was approximately 7 IU of rubella virus-specific IgG per ml of serum sample. For saliva samples the performances of the capture ELISA and previously described radioimmunoassay were assessed, and the results of those two assays were compared to the rubella virus-specific IgG result obtained by a commercial ELISA (Behring Enzygnost) with a panel of paired serum and saliva samples. This comparison showed that the capture ELISA with saliva was more sensitive than the radioimmunoassay and that the results correlated better with the serum IgG result than the results of the radioimmunoassay did, with an overall sensitivity of 82% and a rank correlation of 0.68, whereas the sensitivity and rank correlation for the radioimmunoassay were 74% and 0.45, respectively. For subjects of 10 years of age or younger, the ELISA with saliva had a sensitivity of 94% and a specificity of 100% compared to the results of the ELISA (Behring Enzygnost) for rubella virus-specific IgG with corresponding serum samples. The sensitivity was much lower for subjects ages 17 years or older. The assay may have wider epidemiological use with saliva specimens, particularly those from children.

Effects of the route of infection on immunoglobulin G subclasses and specificity of the reovirus-specific humoral immune response.

Reovirus serotype 1, strain Lang (T1/L), a well characterized enteric virus, elicits humoral and cellular immune responses in mice. Although orally and intradermally induced infections generate comparable reovirus-specific serum antibody titers, little is known about the effects of the route of infection on the systemic immunoglobulin G (IgG) response. To assess whether the route of exposure affects virus-specific humoral immunity, we infected various strains of mice with reovirus T1/L by the oral or intradermal routes. At day 10 following infection, virus-specific serum antibody titers and IgG subclasses were determined by enzyme-linked immunosorbent assay. Serum IgG2a and IgG2b antibodies were detected in all mouse strains independent of the route of infection. Mice of the H-2d haplotype that received an intradermal infection also had high levels of reovirus-specific serum IgG1. This dichotomy of responses was not associated with differences in the types of cytokine produced by draining peripheral lymph nodes. However, peripheral lymph node lymphocytes from C3H mice produced significantly higher levels of gamma interferon than did BALB/c, C57BL/6, and B10.D2 mice. Additionally, peripheral lymph node lymphocytes from all strains of mice produced only low levels of interleukin-5, with no detectable level of interleukin-4 or interleukin-6. Analysis of specific antibody at inductive sites of the immune response showed that orally infected Peyer's patches produced predominantly IgA and intradermally infected peripheral lymph nodes produced predominantly IgG2a. Western blot (immunoblot) analysis showed that virus-specific IgA, IgG1, and IgG2a reacted with reovirus structural proteins. These data suggest that the route of infection affects the isotype and IgG subclasses, but not the antigen specificity, of the local antibody response. In addition, virus-specific IgG1 generated following an intradermally induced infection is linked to the H-2d major histocompatibility complex haplotype.