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The immunogenicity and efficacy of a herpes simplex virus type 2 glycoprotein D (gD2) DNA vaccine formulated with bupivacaine was evaluated using murine and guinea pig models of genital herpes. Animals received three doses of 100 microg of gD2 plasmid or control plasmid intramuscularly prior to intravaginal challenge with HSV-2. Immunization induced HSV ELISA and neutralizing antibody in serum and ELISA antibody in the vaginal secretions of all animals evaluated. Following intravaginal HSV-2 challenge, vaginal viral replication was reduced in both models with peak reductions of greater than 99%. Immunization also decreased the number of animals developing any clinical disease (p < 0.001) and the severity of the acute disease (total lesion score 6.4 versus 0.6 in guinea pigs, p < 0.001). Further recurrent lesion days were reduced from 14.5 to 4.9 days in immunized guinea pigs (p < 0.001). DNA immunization with gD2 + bupivacaine was effective in reducing clinical disease and viral replication in both guinea pigs and mice.

We determined the prevalence of herpes simplex virus (HSV) and cytomegalovirus (CMV) antibodies in a cohort of adolescents 12 to 22 years of age in anticipation of the development of vaccines to control HSV and CMV infections. For the overall study population, we found that 62% were seropositive for HSV type 1 (HSV-1), 12% were seropositive for HSV type 2 (HSV-2), and 65% were seropositive for CMV. Race was not related to HSV-1 seropositivity, but African-American adolescents were more likely than Caucasian adolescents to be seropositive for HSV-2 and CMV. Girls also were more likely than boys to be seropositive for HSV-2 and CMV. For boys, history of a sexually transmitted disease was identified as a risk factor for HSV-2 seropositivity; for girls, a greater number of sexual partners increased the risk of being seropositive for HSV-2. Our data demonstrating a high prevalence of infection during adolescence suggest that immunization for HSV-1, HSV-2, and CMV may need to occur in childhood.

Neonatal herpes simplex virus (HSV) infection produces severe disease with unacceptable morbidity and mortality with current antiviral therapies. The effect of therapy with passive anti-HSV antibody and acyclovir was evaluated using a guinea pig model of neonatal HSV. Newborn animals were inoculated intranasally and treated with acyclovir (60 mg/kg/day) or antibody (or both), beginning on days 0, 2, or 3 after HSV-2 inoculation. Acyclovir alone was effective only when begun on day 0, and antibody alone was effective when begun on or before day 2. Only combination therapy was effective on day 3, reducing mortality from 82% (14/17) in controls to 44% (7/16; P < .05). Combined therapy also significantly reduced the duration of skin, eye, and mouth disease and respiratory symptoms but not recurrent disease. These data suggest that addition of antibody therapy to acyclovir may improve the outcome of neonatal HSV disease.

Cytomegalovirus (CMV) is the most common congenital viral infection, but little is known about the protective immune mechanisms. The guinea pig (gp) model of congenital CMV was used to evaluate the effects of passive antibody given to pregnant dams on pup survival. Dams received three doses of high-titer gpCMV or control antibody on days -3, -1, and +7, or +1, +3, and +7, in relation to gpCMV challenge. gpCMV was inoculated in the late second to early third trimester at three different doses. Compared with controls, gpCMV antibody begun before gpCMV challenge significantly increased pup survival from 14% to 52%, 21% to 84%, and 51% to 77%, respectively, for the three challenge doses. gpCMV antibody started after viral challenge increased pup survival after only the lowest challenge dose (51% to 81%). Antibody did not protect against CMV infection of the pups. CMV antibody appeared to improve survival in congenital CMV infection but did not affect vertical transmission.

In a large placebo-controlled efficacy trial of the rhesus tetravalent (RRV-TV) and serotype 1 monovalent (RRV-S1) rotavirus vaccines in multiple sites throughout the United States, protection against rotavirus disease over a 2-year period was found to be 57 and 40%, respectively (Bernstein et al., J. Am. Med. Assoc., 1995, 273, 1191-1196). Sera collected from a subset of subjects during this trial were used to determine possible correlations between rotavirus antibody responses after vaccination and protection. Between 82% (RRV-S1) and 92% (RRV-TV) of the vaccinees seroconverted by at least one of the six antibody assays performed (i.e. rotavirus IgA and neutralizing antibody to RRV and serotype 1-4 human rotaviruses). Rises in neutralizing antibody were due primarily to RRV. The seroconversion rate was only 18-22% to each of the four human rotavirus serotypes following RRV-TV vaccination and was only 43% to serotype 1 human rotavirus after RRV-S1 administration. Furthermore, no correlate of immunity against rotavirus infection or disease was identifiable based on seroconversion to any of the antibodies measured. Likewise, no consistent relationship was found between the titers of any of these six antibodies following vaccination and protection against rotavirus, thus suggesting that serum antibody titers will not be useful markers of protection with these reassortant RRV vaccines. In addition, vaccinated subjects did not develop higher titers of neutralizing antibody to human rotaviruses following a subsequent natural rotavirus illness, a further indication that only weak immune responses to human rotaviruses were stimulated by vaccination with the RRV reassortants.(ABSTRACT TRUNCATED AT 250 WORDS)

A murine model of genital infection with a thymidine kinase-deficient (tk-) strain of herpes simplex virus type 2 (HSV-2) was utilized to examine the local urogenital antibody response to HSV-2. Vaginal inoculation with HSV-2 tk- protected against a subsequent genital challenge with a lethal dose of virulent HSV-2. After primary vaginal infection, predominantly HSV-specific IgG antibodies were detected in serum and vaginal secretions. HSV-specific IgG antibody-secreting cells (ASC) were detected first and in greatest numbers in the genital lymph nodes (gLN) after primary HSV-2 tk- inoculation. HSV-specific IgG or IgA ASC were not detected in the urogenital mucosa after primary HSV-2 vaginal infection. Vaginal HSV-2 challenge of HSV-immune mice resulted in increased HSV-specific serum IgG antibody and vaginal IgA antibody titers. HSV-specific IgG ASC were detected by 4 days postchallenge in gLN and by Days 6 and 7 postchallenge in the spleen and genital mucosa. These results suggest that urogenital humoral responses originate in the gLN following HSV genital infection and that gLN may serve as the primary source of the HSV-specific IgG- and IgA-secreting cells present in the urogenital mucosa after vaginal challenge.

Determination of protective efficacy after natural rotavirus infection is important as a basis for evaluating rotavirus vaccines. Therefore, placebo recipients in a large 2-year rotavirus vaccine trial conducted across the United States were followed to determine the protection afforded by natural rotavirus infection. Serotype 1 rotaviruses predominated (93% year 1, 66% year 2), but isolates of all four major human rotavirus serotypes circulated during both years. Of the 45 placebo recipients with documented rotavirus illnesses in year 1, 1 developed rotavirus disease in year 2 compared with 29 of the other 235 placebo recipients (P = .03). Serologic data were available for 171 placebo recipients, and 37 of 140 without rotavirus illnesses in year 1 had evidence of asymptomatic rotavirus infection. None of these 37 experienced rotavirus disease in year 2 compared with 22 of the remaining 103 (P < .001). Overall efficacy after natural rotavirus infection was 93% (95% confidence interval, 50%-99%).

Natural infection by very similar strains of rotavirus during the 1988-1989 rotavirus season in Cincinnati, Ohio, provided complete protection of young children against subsequent rotavirus illnesses for a period of at least 2 years. Using this limited strain variability, we characterized the association between the titers of antibody to either the VP4 or the VP7 neutralization protein and protection against subsequent rotavirus disease. This was done by using reassortants that contained only one of the two rotavirus neutralization proteins of 89-12, a culture-adapted isolate representative of the protective rotavirus strains. The other neutralization protein in these reassortants was derived from a heterologous rotavirus (WC3 or EDIM) to which the infected subjects made little or no neutralizing antibody (titers, < or = 20). The geometric mean titer (GMT) of antibody to 89-12 in convalescent-phase sera from the 21 subjects analyzed was 2,323. The GMT of antibody to a reassortant (strain WC-4) that contained the VP7 protein of 89-12 and VP4 of WC3 was 387. In contrast, the GMT of antibody to a reassortant (strain EDIM-7) that contained the VP4 protein of 89-12 and the VP7 protein of EDIM was 1,078. Thus, the major neutralization response was directed against VP4 rather than VP7, a finding that has important implications for development of appropriate rotavirus vaccines.

To determine whether salivary antibody is a reliable indicator of rotavirus infection and mucosal rotavirus antibody concentrations, salivary rotavirus antibody titers were determined as a function of time following inoculation of 24 adult volunteers with a virulent strain of human rotavirus (CJN). Twenty of the subjects became infected and all produced detectable amounts of salivary rotavirus IgA. These antibody concentrations remained undetectable in the four uninfected subjects. Rises of greater than or equal to fourfold in either salivary rotavirus IgA or neutralizing antibody to the challenge virus were detected in all but two subjects. The titers of both antibodies were maximal at 13 days after inoculation and decreased significantly (p less than .02) by 27 days, a result very similar to that previously found with stool rotavirus IgA in CJN-infected subjects. These results suggest that salivary rotavirus antibody titers are an accurate reflection of mucosal rotavirus antibody and a possible surrogate for intestinal rotavirus antibody.

In an evaluation of WC3 bovine rotavirus (serotype 6) vaccine in infants, some subjects experienced a natural serotype 1 rotavirus infection before vaccination and others after. Therefore, the effects of both WC3 and natural rotavirus strains as either primary or boosting immunogens on serotype-specific neutralizing antibody responses could be determined. After primary natural infection (symptomatic or asymptomatic), neutralizing antibody titers were highest to serotype 1 but were consistently high to serotype 3, and low titers (greater than or equal to 20) to serotypes 2 and 4 were often detected. Previous vaccination with WC3 had little effect on the magnitude of these responses. In contrast, subjects infected with serotype 1 strains before vaccination experienced large (average, 12-fold) rises in neutralizing antibody to human serotypes 1-4 when vaccinated with WC3. Thus, although WC3 and the natural strains are distinct serotypes, their epitopes were sufficiently similar that reinfection with WC3 could boost neutralizing antibody titers to human serotypes in subjects primed by a previous natural infection.

The safety, immunogenicity, and efficacy of WC3 rotavirus vaccine was evaluated in a double-blind, placebo-controlled trial of healthy infants 2-12 months of age; 103 received one dose of vaccine and 103 received placebo. Vaccination appeared to be safe and induced an antibody response (WC3 neutralizing antibody) in 97% of vaccinees. Only 9 (9%) of these, however, produced antibody to human rotavirus serotypes; at least 7 of the 9 were naturally infected before vaccination. Neither the number of symptomatic episodes of rotavirus diarrhea (21 vs 25) nor the number of moderate to severe rotavirus illnesses (9 vs. 15) was significantly different in vaccine or placebo recipients, respectively, during a predominantly serotype 1 rotavirus season. A slight but significant decrease in mean symptom score was detected in vaccine recipients. Despite an overall lack of efficacy, protection could be correlated to previous rotavirus infection, high levels of WC3 neutralizing antibody, and preexisting (maternal) serotype 1 neutralizing antibody with a titer greater than or equal to 30.

The immune response to guinea pig cytomegalovirus (gpCMV) was evaluated by immunoblotting. Preinoculation guinea pig plasma did not react with gpCMV antigen, whereas convalescent plasma reacted to at least 18 gpCMV-specific polypeptides. The initial immune response was primarily directed at polypeptides with MWs of 100, 75, and 56 kDa. Over 80% of plasma collected more than 29 days after viral inoculation reacted to these polypeptides and also to those with MW of 54, 52, and 38 kDa. In this report, we also demonstrate cross reactivity between gpCMV and human CMV (HCMV). Human immunoglobulin (IVIG) reacted to at least 20 HCMV polypeptides and cross reacted with six gpCMV polypeptides. GpCMV convalescent plasma also reacted with HCMV polypeptides.

Serum specimens from infants 2 to 12 months old vaccinated with the WC3 bovine rotavirus were analyzed to determine the relative concentrations of neutralizing antibody to the VP4 and VP7 proteins of the vaccine virus. To do this, reassortant rotaviruses that contained the WC3 genome segment for only one of these two neutralization proteins were made. The segment for the other neutralization protein in these reassortants was from heterotypic rotaviruses that were serotypically distinct from WC3. Sera were examined from 31 infants who had no evidence of a previous rotavirus infection and the highest postvaccination WC3-neutralizing antibody titers (i.e., 160 to 600) of the 103 subjects administered the vaccine. A reassortant (3/17) that contained both neutralization proteins from the heterotypic rotaviruses, i.e., EDIM (EW strain of mouse rotavirus) VP7 and rhesus rotavirus VP4, was not neutralized by these sera (geometric mean titer [GMT], less than 20). A reassortant (E19) that contained EDIM VP7 and WC3 VP4 was also very poorly neutralized by these antisera (GMT = 20). In contrast, antibody titers to a reassortant (R20) that contained WC3 VP7 and rhesus rotavirus VP4 were higher than those against WC3 (GMTs of 458 and 313, respectively). Thus, VP7 appeared to be the dominant immunogen for production of neutralizing antibody after intestinal infection of previously uninfected infants vaccinated with WC3 bovine rotavirus.

Previous studies of adults challenged with human rotavirus (CJN strain) showed that 74% became infected and 55% of those infected experienced illness. Protection against infection correlated with rotavirus antibody, most significantly (P = .005) serum rotavirus IgG. In this study, 20 previously challenged subjects were reinoculated with the same virus 9-12 months after their initial challenge. Only 1 of 8 subjects not infected after the initial challenge and 2 of 12 infected after the first inoculation became infected after reinoculation; none became ill. Titers of rotavirus antibodies (serum, jejunal, and stool) at the time of reinoculation were about as high as or higher than they were before the initial inoculation. This correlated with greater protection, but the extent of protection was significantly greater (P less than .0001) than predicted based on a previous model relating protection and preinoculation titers of serum rotavirus IgG. Thus, inoculation with human rotavirus provided homotypic protection for at least 9-12 months, and protection remained correlated with higher concentrations of rotavirus antibody. However, the specific relationship between protection and rotavirus antibody was altered after the initial inoculation.

The antibody response to herpes simplex virus (HSV) glycoproteins B and D was evaluated using these cloned glycoproteins in an ELISA assay and compared to a standard Western blotting procedure. The ELISA assay appeared to be more sensitive for detecting gB and gD antibodies. Antibodies to gB but not gD were detected in the acute sera of patients presenting with their first episode of true primary genital herpes. The geometric mean HSV gB titer (log 2) was also significantly higher than the geometric mean gD titer in the convalescent sera of these patients (7.9 +/- 0.7 vs. 5.5 +/- 0.9, P less than .003). A cross-reaction between HSV gB and varicella zoster virus (VZV) gp II was demonstrated. Thus, it is possible that a previous VZV infection could prime the immune system to respond rapidly and more vigorously to HSV gB. Indeed, in this report we demonstrated a significant correlation between the VZV ELISA absorbance and the titer to HSV gB and also detected a higher VZV ELISA absorbance and HSV gB titer in the acute sera of patients with a true primary HSV infection compared to other HSV seronegative VZV seropositive patients. Use of this quantitative assay should allow further investigation into the relationship of the immune response to these important targets and the clinical course of HSV disease.

The humoral response to herpes simplex virus (HSV) was examined in 21 patients with primary HSV infection who were participating in a placebo-controlled trial of oral acyclovir as therapy for first-episode genital herpes. A standard microneutralization assay and a western blot analysis, which detects the antibody response to individual polypeptides of HSV types 1 and 2, were used. Placebo-treated, HSV type-2-infected patients typically responded to HSV type-2 polypeptides that had molecular masses of 125, 84, 68, 59, 46, and 40 kilodaltons. Treatment with acyclovir diminished the humoral response both to HSV, as measured by the type-specific neutralizing-antibody titer, and to specific polypeptides, especially those with molecular masses of 50-100 kilodaltons, a range that includes polypeptides with molecular masses corresponding to those of glycoproteins D and E of HSV.

The production of antibody to specific herpes simplex virus type 2 (HSV-2) polypeptides, the recurrence patterns and the susceptibility to re-infection were studied in the guinea-pig model of genital HSV-2 infection. Further, we defined the effects of acyclovir (ACV) therapy on these parameters of infection. Treatment with ACV reduced the clinical severity of the initial disease but did not affect vaginal viral shedding. Production of neutralizing antibody as well as antibody to the nucleocapsid protein, and glycoproteins B, D and G were all delayed in ACV recipients. ACV treatment of initial infection did not significantly alter the pattern of subsequent recurrence although by several criteria treated animals tended towards decreased recurrences. Re-inoculation with a second strain of HSV-2 resulted in a local cervicovaginal infection but, except for one ACV-treated animal, neural tissue was protected from re-infection.

The serological response to the ICP-35 group of herpes simplex virus type 2 (HSV-2) proteins was examined 30 d after vaginal challenge with HSV-2 of guinea pigs previously immunized with HSV glycoproteins. Detection of antibodies to ICP-35 proteins correlated best with the magnitude of vaginal viral replication. Animals that developed antibody to ICP-35 proteins had more-frequent recurrences (P less than .0002) that were of longer duration (P less than .005) than animals that did not. By covariate analysis the number of recurrences appeared to be related to the severity of the acute disease, inversely related to the enzyme-linked immunosorbent assay titer of antibody to HSV glycoproteins, and related to the detection of antibody to ICP-35 proteins. The duration of recurrent disease, however, could only be correlated with the presence of antibody to ICP-35 proteins. Detection of antibody to ICP-35 proteins is thus a useful prognostic indicator for identifying animals that developed more-severe recurrent disease.

Anti-influenza serum and nasal antibody titers were followed during a double-blind, placebo-controlled, randomized study assessing the prophylactic efficacy of 50, 100, and 200 mg/day of amantadine hydrochloride against experimental challenge with influenza A/Beth/1/85. The geometric mean titers (GMT) of serum hemagglutination inhibition antibody (P = .05) as well as serum influenza-specific IgG (P = .004) were significantly lower for all amantadine groups when compared to placebo. There were no significant differences in the GMT of either type of serum antibody titer comparing any of the three amantadine groups. Viral titers were also lower in the amantadine groups compared to the placebo group and not significantly different among the three amantadine groups. Nasal antibody titers in the 100-mg and 200-mg amantadine groups were significantly lower than placebo titers (P = .002). Nasal antibody titers in the 50-mg amantadine group did not differ significantly from titers in the placebo group (P = .892). Possible reasons for the differential effect of amantadine on the serum and nasal antibody response are discussed.

Because rubella continues to be a common illness in adolescents and young adults and because it has been suggested that booster rubella immunizations should be performed, we studied antibody prevalence in 459 predominantly adolescent patients in a pediatric group practice. Rubella antibody (PHA titer greater than or equal to 1:13.5) in previously immunized patients (89.6% of 385) was significantly more common than antibody in unimmunized patients and patients with a questionable history of immunization (70.3% of 74) (P less than 0.005). Twenty-three seronegative patients with a documented history of prior immunization were reimmunized and 22 had an IgG (secondary) antibody response and only one an IgM (primary) antibody response. Since all but one of our patients with previous immunization had a secondary immune response following revaccination, it seems likely that the level of protection in previously vaccinated individuals is considerably greater than 90%. Attention today should be directed at finding and immunizing unvaccinated teenagers and young adults and not in major booster vaccine programs.

Acute serum samples from patients with nonprimary first-episode genital herpes were evaluated for the presence of type-specific antibody to herpes simplex virus type 1 and type 2. A standard microneutralization assay and western blot analysis, which identified the response to individual polypeptides of type 1 and type 2 herpes, were performed in both unadsorbed serum samples and serum samples adsorbed with either type 1 or type 2 antigens to remove cross-reacting antibodies. Of 24 serum samples examined, seven were found to contain only type 1 antibodies, 11 had type 2 antibodies alone, and six serum samples had both type 1 and type 2 antibodies. It is believed that a majority of the 17 patients whose acute serum samples already contained type 2 antibody were experiencing their first clinically apparent recurrence of a previous asymptomatic type 2 infection. Physicians should, therefore, acknowledge that patients with first-episode nonprimary herpes 2 infection may not have had a recent sexual contact with a herpes simplex virus-infected partner.

Effects of preinoculation rotavirus antibody titers on the probability of infection and illness were evaluated in adults challenged orally with different doses of a virulent human rotavirus (CJN strain). Preinoculation titers considered were serum neutralizing antibody, serum rotavirus IgA, serum rotavirus IgG, jejunal neutralizing antibody, jejunal rotavirus IgA, and stool rotavirus IgA. Doses of virus of either 9 x 10(1) or 9 x 10(3) focus-forming units were administered to 19 subjects each. Twenty-six were infected; 15 experienced illness. The probability of either outcome was unrelated to dose. Stool rotavirus IgA titers could not be correlated to either infection or illness, but the mean titers of the other five antibodies were significantly or nearly significantly lower in subjects infected or ill, when compared with those negative for either outcome. When analyzed by stepwise logistic regression, only serum rotavirus IgG remained significantly (P = .005) related to the probability of infection, and only jejunal neutralizing antibody remained significantly (P = .01) related to the probability of illness.

The reactogenicity and immunogenicity of a trivalent split-product influenza vaccine and a trivalent subunit influenza vaccine were studied in 67 children and young adults. Subjects received two doses of either vaccine given one month apart. The reactogenicity of the two vaccines was similar. After the second dose of the split-product vaccine, the prevalence of hemagglutination-inhibiting (HAI) antibody titers of 20 or more was as follows: A/Brazil/78, 73%; A/Bangkok/79, 100%; and B/Singapore/79, 73%. Similar data for the subunit vaccine recipients were 78%, 100%, and 74%, respectively. The HAI antibody responses to the A/Brazil/78 and B/Singapore/79 antigens of both vaccines were relatively poor in those subjects who were initially seronegative (HAI antibody titer less than 5) to the specific antigen.

Herpes simplex virus type 1 (HSV1) and type 2 (HSV2) polypeptides with type-common and type-unique epitopes were identified using cross-adsorbed hyperimmune rabbit sera and Western blotting techniques. Twelve HSV1 and fourteen HSV2 polypeptides with type-specific epitopes were identified. Cross-adsorbed human sera reacted to a subset of the type-specific epitopes defined by rabbit sera. Human sera with both HSV1- and HSV2-specific antibodies were identified by their reaction to HSV1-type-specific epitopes after adsorption with HSV2, and HSV2-type-specific epitopes after adsorption with HSV1. Mixing experiments demonstrated the sensitivity of this assay for detecting HSV2-type-specific antibody in the presence of HSV1 antibody. This technique has proven useful in the study of immune response to individual HSV polypeptides during human HSV infections, while the presence of these type-unique epitopes on HSV polypeptides allows for the detection of type-specific antibodies in human sera.

We developed an enzyme-linked immunosorbent assay (ELISA) for measuring the antibody response to herpes simplex virus (HSV) glycoprotein D (gD). The ELISA was specific and more sensitive than immunoblotting techniques. Antibody to HSV gD was detected in 10 of 12 sera (geometric mean titer, 36.3) obtained from HSV-2-infected guinea pigs 14 days after intravaginal inoculation, and the titer increased to greater than or equal to 5,120 on day 60. Therapy with acyclovir delayed and diminished the antibody response to gD, although by day 60 the titers of antibody to gD in acyclovir-treated animals were not significantly different from those in controls. An increased titer of antibody to gD seemed to be associated with a reduced number of recurrent episodes and to a reduced number of days with recurrent lesions.

Peripheral blood leukocytes (PBL) from 18 homosexual men who did not have acquired immunodeficiency syndrome (AIDS) and from 9 heterosexual men were repetitively tested for their ability to generate HLA self-restricted cytotoxic T lymphocyte responses to influenza virus (flu-self) over a 2-yr period. The sera of the same donors were tested for antibodies to human T lymphotropic virus-III (HTLV-III). Six of the homosexual and none of the heterosexual donors consistently generated weak cytotoxic T lymphocyte responses to flu-self. Seven of the homosexual and none of the heterosexual donors were seropositive for antibodies to HTLV-III. No obvious correlation was detected between weak flu-self cytotoxic T lymphocyte responses and antibodies to HTLV-III. However, one homosexual donor generated no detectable cytotoxic T lymphocyte activity to flu-self, although he was a strong responder to HLA-alloantigens. This donor had an OKT4:OKT8 ratio of 0.4 and was seropositive for HTLV-III antigens; HTLV-III virus was identified in his PBL; and he developed AIDS during the course of this study. A second donor with lymphadenopathy and who was seropositive for HTLV-III antigens exhibited marginal cytotoxic T lymphocyte activity to flu-self which he subsequently lost. PBL from two patients, one with Kaposi's sarcoma and one with generalized lymphadenopathy, were also tested for cytotoxic T lymphocyte responses to flu-self and to alloantigens. Both donors failed to generate cytotoxic T lymphocyte to flu-self, but generated strong cytotoxic T lymphocyte responses to alloantigens. The selective loss of an HLA-restricted cytotoxic T lymphocyte response without loss of HLA alloantigenic cytotoxic T lymphocyte activity may be an important functional immunologic characteristic in the development of AIDS.

The induction and persistence of local rotavirus antibodies, including stool IgA, jejunal IgA, and jejunal neutralizing antibody, were evaluated in 14 adult volunteers infected with the CJN strain of human rotavirus. In addition, the relationships between local rotavirus IgA and serum rotavirus IgA, IgG, and neutralizing antibody were determined. Both stool and serum rotavirus IgA appeared to have similar kinetics. Both antibodies peaked by days 14-17 after inoculation in all subjects, then decreased rapidly. By days 26-28, titers had fallen to 13% and 30% of their respective peaks. Serum rotavirus IgG peaked somewhat later, occurring in five subjects on days 26-28. Serum neutralizing antibody peaked on days 26-28 in all but three subjects. Both serum IgG and neutralizing antibodies also declined more slowly than rotavirus IgA. Although all antibody concentrations had decreased to only a fraction of their peak responses by days 270-365 after rotavirus inoculation they remained higher than baseline levels. For example, stool rotavirus IgA concentrations were 13.5-fold higher than baseline, while jejunal rotavirus IgA and neutralizing antibody were 8.9- and 4.3-fold above baseline, respectively. Similarly, serum antibodies remained 3.7- to 11.2-fold higher than baseline at 270-365 days after rotavirus inoculation. These studies imply that serum rotavirus IgA is a good indicator of local antibody responses. Furthermore, although both serum and local antibody titers peaked within 2-4 weeks after infection, these antibodies persisted at above baseline concentrations for at least 9-12 months after infection.

A newly developed system (as described in the previous paper) employing separated herpes simplex virus (HSV) polypeptides electrophoretically transferred to nitrocellulose paper, Western Blot Analysis (WBA), was compared to microneutralization (MN) for the detection of type-specific antibody to HSV types 1 and 2 in 98 human sera. Sera containing HSV1-specific antibodies reacted at least twofold more densely (as quantitated by densitometric scanning) to HSV1 than HSV2 polypeptides, while sera with HSV2-specific antibodies reacted twofold more densely to HSV2 polypeptides. The reliability of this system was determined (1) by analysis of sera from patients with true primary herpes simplex infections, (2) by adsorption studies that removed cross-reacting antibodies, and (3) by comparison with neutralization. Of 36 sera found to have HSV1-type-specific antibody by WBA, 83% had MN ratios of less than 85 (indicating HSV1 antibody), while of 28 sera found to have HSV2-type-specific antibody by WBA, 89% had MN ratios of greater than 85 (indicating HSV2 antibodies). The MN ratio obtained on sera felt to have both HSV1-type-specific and HSV2-type-specific antibody by WBA ranged from 67 to 160. WBA agreed more closely with clinical isolates and adsorption studies than MN. This system not only accurately determines the HSV-type-specific antibody status of patients but provides important information on the specific immunogenic proteins.

An evaluation of the safety and immunogenicity of WC3 rotavirus vaccine was evaluated in adult volunteers. Pre- and post-vaccination titers of neutralizing antibody to WC3 and to the four human rotavirus serotypes as well as serum and stool rotavirus IgA levels were measured. Vaccination was safe and did not induce elevation of liver enzymes. None of the 12 volunteers receiving WC3 vaccine shed detectable amounts of virus although antibody rises were detected in 11 of 12 vaccines. Nine developed and increase in WC3 neutralizing antibody, one additional subject had a rise in Wa (human serotype 1) neutralizing antibody while another subject only developed a rise in stool rotavirus IgA. All of the vaccine recipients with a rise in WC3 neutralizing antibody also developed a rise in neutralizing antibody against at least one of the four most common human rotavirus serotypes. A stool IgA rotavirus antibody response was detected in 6 of 9 WC3 recipients with measurable stool antibody. None of the control subjects developed significant rises in any of the antibody titers measured. WC3 rotavirus vaccine appears to be safe and induces systemic and local immune responses in adults suggesting that further evaluation of WC3 should be considered in infants.

Our studies of rotavirus challenge in adult volunteers enabled us to evaluate the relationship of pre-existing antirotavirus fecal IgA antibody to infection and illness and to investigate the local response to this infection. No relationship could be found between the pre-existing levels of fecal antirotavirus IgA antibody and protection from infection or illness. A greater than six-fold increase in the level of antibody was seen in 16/19 infected volunteers with determinable increases but in 0/15 controls who received less than the minimal infectious dose of rotavirus. Antibody levels increased rapidly in infected volunteers and were consistent with an anamnestic response. Two of seven volunteers who received an infectious dose of rotavirus but were considered uninfected on the basis of other laboratory methods had greater than or equal to six-fold increases of fecal antibody and one of these experienced symptoms compatible with a rotavirus infection. This finding indicates that an increase in fecal antibody may be a reliable indicator of rotavirus infection even in the absence of detectable shedding or seroconversion.