Primary Genital Infection Research Articles

Indirect ELISA for the detection of HSV-2 specific IgG and IgM antibodies with glycoprotein G (gG-2)

The glycoprotein G (gG-2) purified from HSV-2 infected cells has been reported to be useful for determination of HSV-2 type-specific antibodies using conventional ELISA formats. This study further confirmed the specificity of gG-2 and demonstrated the feasibility of a specific IgM assay. The gG-2 ELISA was developed to detect HSV-2 specific IgG and IgM antibodies in human sera with high levels of sensitivity and specificity. Of 45 patients with culture-proven recurrent HSV-2 genital infection 44 were reactive for gG-2 IgG. Of 30 sera from patients with culture-proven recent initial HSV-2 genital infection 29 were positive for gG-2 IgM. Three patients with primary HSV-2 genital infection showed gG-2 IgM in the convalescent but not in the acute sera. The IgG- and IgM-gG-2 ELISA showed high specificity. None of 40 sera from children were reactive by either assay. Only one of 94 sera from patients with antibody to herpesviruses other than HSV reacted in the IgG assay but none reacted in the IgM assay. There was no cross-reaction with sera from patients with proven HSV-1 infection with the gG-2 antigen. The results suggest that the IgG assay can be used for demonstration of past HSV-2 infection and the IgM assay for the diagnosis of HSV-2 in neonatal herpes and primary genital herpes, when cultures or rapid diagnostic techniques are unavailable.

NEW ASPECTS OF NEONATAL HERPES

Herpes simplex virus (HSV) is a rare but increasingly prevalent agent of newborn infection. Primary or first episode genital infection in the mother constitutes the greatest risk of infection in the neonate. Transplacental, HSV-specific functional antibody appears to protect infants from severe disease and may be therapeutically exploitable. Prevention of serious sequelae in the infant depends on early diagnosis and treatment of HSV-infected infants.

Vaginal infection of mice with HSV type 2 variant ER −: A new animal model for human primary genital HSV type 2 infections

Studying the pathogenesis of vaginal infections in mice with two variants of Herpes simplex virus type 2 (HSV-2) strain ER we observed that both variants ER + and ER − caused severe vaginitis but only ER + invaded the CNS leading to lethal neurological disease. In contrast, mice infected with ER − cleared the virus from the vagina and recovered from infection. ER + and ER − expressed equal levels of thymidine kinase (TK) indicating a TK-independent difference in neurovirulence. Using the non-neurovirulent variant ER −, we were able to investigate humoral immune responses late after infection. Vaginal infection with ER − suppressed serum antibody formation after a secondary systemic HSV-1 infection. Fresh isolates of HSV-1 and HSV-2 caused uniformly a lethal neurological disease after vaginal inoculation of mice. However, some animals survived an intraperitoneal infection with these isolates. Infection with HSV-1 isolates stimulated a strong antibody production, whereas infection with HSV-2 isolates suppressed antibody formation, thus supporting earlier results from our group obtained with laboratory strains. Since suppression of antibody formation could be demonstrated with clinical HSV-2 isolates and likewise after vaginal infection with HSV-2 variant ER −we consider this phenomenon to be of relevance in human genital HSV-2 infections. Vaginal infection of mice with variant ER − represents a new model for primary genital HSV-2 infections; this model could be useful for histopathological, virological, immunological and drug testing studies.

Susceptibility to reinfection after a primary chlamydial genital infection is associated with a decrease of antigen-specific T cells in the genital tract

We tested the hypothesis that the intensity of specific antichlamydial T cell-mediated immunity in the genital tract of female guinea pigs infected intravaginally with the chlamydial agent of guinea pig inclusion conjunctivitis would determine the resistance or susceptibility to reinfection after a primary chlamydial infection. T cell-enriched lymphocytes were isolated by collagenase treatment of genital tract tissues from either infected or control uninfected female guinea pigs at various times after infection. The nylon wool-enriched T lymphocytes were evaluated for expression of antigen-specific T cell-mediated immunity in vitro by using a blast transformation assay. Both uninfected and infected genital tracts contained T cells, as evidenced by reactivity to concanavalin A, although a greater number of T lymphocytes was detected in the genital tracts of infected animals compared with that in controls. Significant antigen-specific T-cell activity could be detected in the genital tract tissue by 7 days after a primary genital tract infection with the chlamydial agent of guinea pig inclusion conjunctivitis. When antigen-specific activity was assessed at different times after infection, the intensity of the response of genital tract-associated T lymphocytes was directly proportional to the degree of resistance of the animals to genital challenge. Thus, susceptibility of animals to reinfection by chlamydiae appears to be associated with the intensity of the local T cell-mediated immune responses in the genital tract of infected animals.

Capsaicin Interferes with the Centrifugal Spread of Virus in Primary and Recurrent Genital Herpes Simplex Virus Infections

The neural spread of herpes simplex virus (HSV) via sensory nerves is an essential component of disease pathogenesis. Capsaicin, a specific toxin for sensory C-type fibers, interferes with neuronal function including axonal transport. Capsaicin treatment of guinea pigs before intravaginal HSV inoculation did not prevent ganglionic infection but ameliorated cutaneous herpetic disease, suggesting that anterograde but not retrograde virus spread involved a capsaicin-sensitive process. Similarly, treatment of latently infected animals reduced both spontaneous and ultraviolet radiation-induced herpetic recurrences, suggesting that capsaicin interfered with the anterograde spread of reactivated virus. These results suggest that the centrifugal spread of virus may involve either different pathways or mechanisms than are involved in centripetal HSV spread. The use of selective neuropharmacologic agents such as capsaicin may prove useful in further defining the pathophysiology of HSV disease.

Treatment of Primary and Recurrent Herpes Simplex Virus Type 2 Induced Meningitis with Acyclovir

We describe therapy with acyclovir in 1 patient with acute meningitis induced by herpes simplex virus type 2 (HSV-2) and in 1 patient with ascending myelitis in connection with meningitis after a primary genital HSV-2 infection. In addition, intermittent or continuous acyclovir prophylaxis against meningitis was employed in 3 patients with recurrent meningitis of probable HSV-2 etiology. Possible beneficial effects of the treatment were seen.

Cytology of primary versus secondary genital herpesvirus infection

Certain authors state that primary genital herpes infections mainly produce cells with ground-glass nuclei, whereas secondary infections produce a higher percentage of multinucleated or single cells with prominent intranuclear inclusions. The cervicovaginal smears of 11 patients with primary genital herpes infection and 16 patients with secondary infection were studied to determine the validity of this differentiation. The percentage per patient of virally infected cells containing distinct inclusions ranged from 0% to 35% in the primary herpes group and 0% to 100% in the secondary herpes group. In eight additional patients, the initial herpes simplex virus (HSV)-positive smear was compared with recurrent positive smears taken from 2 to 120 mo later. No statistical difference in the incidence of distinct inclusions in the infected cells was detected. Finally, primary rabbit kidney and the heteroploid A549 cell cultures were inoculated with herpes viruses and followed by fixation with several agents at various time intervals. Alcohol and Bouin's fixative favored the recognition of distinct inclusions, as did the duration of the infection.

Susceptibility to reinfection after a primary chlamydial genital infection

Female guinea pigs which had been infected genitally with the agent of guinea pig inclusion conjunctivitis were challenged at various times after infection with fresh inocula to determine the duration of immunity resulting from the primary infection. At 30 days after infection, most guinea pigs were resistant to reinfection, as indicated by the inability to isolate chlamydiae from cervical swabs. However, at 77, 155, and 294 days, all animals became reinfected, although the course of the infection was abbreviated and of lower intensity. When various immune parameters were examined, a decrease in antibodies in both serum (immunoglobulin G [( IgG]) and genital secretions (IgA, IgG) was observed after 30 days. A decrease in antibodies to the major outer membrane protein and an 84K component was noted in serum. In genital secretions, IgA antibodies to all major chlamydial components declined markedly after 30 days. Cell-mediated immunity as measured by proliferation of peripheral blood lymphocytes to guinea pig inclusion conjunctivitis antigen also was at a peak response 30 days after infection and decreased thereafter. Thus, loss of complete immunity could not be associated with a particular immune parameter. When genital secretions were examined 14 days after the challenge infection, IgA antibody levels to the lipopolysaccharide and 61K protein components had increased in intensity, whereas other antibodies were relatively low. In addition, complete immunity to a third infection was not increased in duration when animals had recovered from two previous genital infections.

Efficacy of Recombinant Glycoprotein D Subunit Vaccines on the Development of Primary, Recurrent, and Latent Genital Infections with Herpes Simplex Virus Type 2 in Guinea Pigs

Vaccination of guinea pigs with recombinant glycoprotein D (subunit vaccine) from either herpes simplex virus type 1 or type 2 (HSV-1, HSV-2), before intravaginal inoculation with HSV-2, markedly altered viral replication in the vaginal tract and reduced, or completely prevented, the development and spread of external genital lesions. Vaccination with glycoprotein D from HSV-2 prevented the establishment of latent HSV-2 infections in dorsal root ganglia and reduced the frequency of recurrent episodes. For all parameters of efficacy that we tested, vaccination with glycoprotein D from HSV-2 was more effective than was vaccination with protein derived from HSV-1. Vaccination after primary infection had no effect on the frequency or duration of recurrent HSV-2 infections.

Comparison of Western blot (immunoblot) and glycoprotein G-specific immunodot enzyme assay for detecting antibodies to herpes simplex virus types 1 and 2 in human sera

Sera from patients with culture-proven genital herpes infections were tested for herpes simplex virus type 1 (HSV-1)- and HSV-2-specific antibodies by both a Western blot (immunoblot) technique (WBA) and immunodot enzyme assays (IEAs) specific for HSV-1 or HSV-2 glycoprotein G (gG). Of 137 serum samples tested, none was mistyped by either WBA or IEA. Both tests were most sensitive with sera obtained at least 21 days after onset of primary HSV-2 infections or sera drawn during recurrent HSV-2 genital episodes: 75 of 76 (99%) such serum samples were positive for HSV-2 antibody by WBA and 73 of 76 (96%) were positive by IEA. Of sera drawn earlier than 21 days from onset of primary genital HSV-2, antibodies to HSV-2 were detected in 25% by WBA and 8% by IEA. In patients with culture-proven primary genital HSV-1 infection, WBA detected antibodies to HSV-1 proteins in 16 of 17 (94%) serum samples drawn at least 21 days after onset of primary genital HSV-1 infection, compared with 9 of 17 (53%) serum samples tested for gG-1 by IEA. Both WBA and IEA are accurate and sensitive tests for HSV-2 antibody in patients convalescing from a first episode or having symptomatic or asymptomatic recurrent genital herpes. WBA was more sensitive than IEA in detecting seroconversion following primary HSV-1 genital herpes, although both assays may miss persons undergoing early seroconversion to HSV-2.

Disseminated Herpes Simplex Infection in a Neonate During Prophylaxis With Vidarabine

HERPES simplex virus (HSV) can cause overwhelming neonatal infection even when antiviral therapy is initiated at the first sign of illness. 1-3 Thus, antiviral prophylaxis has been recommended for high-risk neonates, 1,3 including those delivered vaginally or by See also pp 1051,1056, and 1067. cesarean section (after membranes have been ruptured for more than four hours) to mothers with primary HSV genital infections. 4 This case report describes a high-risk neonate who developed disseminated HSV infection despite antiviral prophylaxis. Report of a Case A 24-year-old woman presented to the hospital 24 hours after rupture of membranes. She was at term and had been followed up for 32 weeks in the obstetrics clinic. She had had intercourse seven days before rupture of membranes. She had no history of HSV infection and no known exposure to HSV. On admission, her only complaint was vaginal pain and burning of two days' duration. Physical

Antiviral treatment of herpes simplex infection in neonates and pregnant women

Herpes simplex infections in neonates include (1) infections beginning in skin or mucous membranes, (2) infections of the central nervous system, and (3) disseminated herpes of the newborn. Early recognition of infections beginning in skin or mucous membranes is essential because 75% will disseminate internally if untreated. Skin lesions provide important diagnostic clues in herpes of the nervous system and disseminated herpes, but they often appear late and in only about 60% of infected neonates. Acyclovir and vidarabine are effective therapies but they must be administered early to prevent serious damage or death. Herpetic skin lesions in neonates may be confused with pyodermas. Direct immunofluorescence and cultures are reliable diagnostic tests. Congenital herpes may cause widespread skin involvement and multiple eye and nervous system diseases. The mother is the source of neonatal herpes in about two thirds of cases. Asymptomatic genital shedding of virus late in pregnancy does not accurately predict whether maternal cultures will be positive at the time of delivery. The risk of the infant acquiring neonatal herpes from a mother with recurrent herpes at birth is about 5%, but the risk is much higher if the mother has true primary genital infection.

Analysis of the humoral immune response to chlamydial genital infection in guinea pigs.

Studies using the guinea pig model of chlamydial genital infection with the Chlamydia psittaci agent of guinea pig inclusion conjunctivitis (GPIC) have shown that serum and local antibodies play a role both in the resolution of infection and in protection against reinfection. Thus, this model is suited for further exploration of immune mechanisms and for vaccine studies with chlamydial macromolecules. We have further characterized the model by assessing the antigen-specific antibody response to experimental genital infection by using immunoblotting to assay both genital secretions and serum. The GPIC agent was characterized by analysis of outer membrane proteins, which indicated that the GPIC agent possessed a major outer membrane protein (MOMP), with a molecular mass of 39 kilodaltons (kDa), and a 61-kDa protein, analogous to cysteine-rich 60-kDa proteins or doublets of Chlamydia trachomatis strains. As indicated by immunoblotting, most infected animals produced serum immunoglobulin G antibodies to MOMP, the 61-kDa proteins, an 84-kDa outer membrane protein, and lipopolysaccharide. Such serum antibodies persisted for at least 813 days after primary genital infection. Immunoglobulin A antibodies against the 61-kDa proteins, lipopolysaccharide, and MOMP, but not the 84-kDa protein, were detected in secretions. Animals challenged with GPIC 825 days after primary infection became infected again despite the presence of serum antibodies, but the period of chlamydial shedding was significantly shorter and less intense than in primary infections. Although the specific mechanism is not known, these data suggest that a long-lasting immune effect is capable of altering the course of infection late after primary infection. Correlation of the antigen-specific antibody response and other immune parameters with the duration and degree of protective immunity induced by infection or vaccination may be helpful in further understanding the nature of such protective immunity.

Intrauterine herpes simplex virus infections

Neonatal herpes simplex virus (HSV) infection is usually acquired at birth, although a few infants have had findings suggestive of intrauterine infection. We describe 13 babies who had clinical manifestations of intrauterine HSV infection, including skin lesions and scars at birth (12), chorioretinitis (eight), microcephaly (seven), hydranencephaly (five), and microphthalmia (two). All infants had combinations of these defects. Infection was proved by viral isolation in each case; all isolates were HSV-2. Two infants died during the first week of life; 10 of the surviving infants had severe neurologic sequelae, and one infant was blind. Four mothers experienced an apparent primary genital HSV infection, and one had recurrent infection, at varying times during gestation. The remaining women denied a history of symptoms of genital HSV infection. These findings indicate that intrauterine HSV infection can occur as a consequence of either primary or recurrent maternal infection and has severe consequences for the fetus.

A novel glycoprotein for detection of herpes simplex virus type 1-specific antibodies

A novel herpes simplex virus type 1 (HSV-1)-specific glycoprotein reactive with monoclonal antibody H1379 was purified by affinity chromatography. This glycoprotein, provisionally designated as gG-1, forms two sets of bands with molecular weights of 40–44,000 and 60–88,000. When used in an immunodot enzymatic assay, gG-1 reacted strongly with rabbit antisera to HSV-1, but not with sera hyperimmune to HSV-2. Specificity of the assay was further established by the lack of reactivity of convalescent sera collected from 20 patients with primary genital HSV-2 infections, and from 100 seronegative individuals. In contrast, antibodies to gG-1 were detected in 9 of 10 patients with primary HSV-1 infection, and in 63 67 patients with culture-positive, recurrent oral or genital HSV-1 infection. Reproducibility of the gG-1 immunodot assay for HSV-1 antibody detection was 96%. Serological assay with purified gG-1, done in parallel with the assay using purified gG-2 described in an earlier report, provides simple and reliable methods to detect type-specific HSV-1 and HSV-2 antibodies for sero-epidemiological studies.

Detection of herpes simplex virus type 2-specific antibody with glycoprotein G.

A recently described herpes simplex virus (HSV) type 2 (HSV-2)-specific glycoprotein (gG-2) was purified on an immunoaffinity column prepared with monoclonal antibody. This purified antigen was used in an immunodot enzymatic assay on nitrocellulose paper for the detection of HSV-2 antibodies in human serum. The test was very sensitive in that HSV-2 antibodies were detected in the convalescent sera of 132 of 134 patients with recurrent genital infections in which HSV-2 had been isolated earlier. Antibodies to gG-2 were detected in 17% of sera obtained within 10 days after the onset of a primary HSV infection and in 95% of sera obtained more than 10 days after onset. The specificity of the immunodot assay was demonstrated by testing sera from 245 HSV-seronegative adults, 344 children, 29 nuns, and 13 patients with primary genital HSV-1 infections. None of these 631 sera was reactive with the gG-2 antigen. When compared with a microneutralization test, the immunodot assay was found to be more specific in detecting HSV-2 antibodies. Reproducibility of the gG-2 assay, obtained by retesting 391 sera, was 95%. Thus, this assay has the sensitivity, specificity, and reproducibility necessary for the measurement of HSV-2 antibodies in seroepidemiological studies.

IgG subclass antibodies to herpes simplex virus.

Several mouse monoclonal antibodies specific for human IgG1, IgG2, IgG3, and IgG4 were evaluated by enzyme-linked immunosorbent assay to detect human IgG subclass antibodies to herpes simplex virus (HSV) antigens. The variable results with different monoclonal antibodies point to the need for well-characterized reagents in the study of antibody responses to infectious agents. The 204 sera tested were obtained from 157 patients with various forms of clinically manifest HSV infections and from several controls. IgG1 antibodies were demonstrated in almost all HSV-infected subjects and were the first antibodies to appear in primary genital infections. IgG2, IgG3, and IgG4 antibodies were detected in acute-phase sera, most often in patients with recurrent genital herpes but in none of those with primary infections. IgG4 antibodies occurred significantly more frequently in sera from men than in those from women with recurrent genital infections.

1114 CONGENITAL HERPES SIMPLEX VIRUS (HSV) INFECTION - EARLY VERSUS LATE GESTATIONAL ACQUISITION

Neonatal HSV infection is usually acquired at birth but a few reports have implied intrauterine infection. Of 155 cases of neonatal herpes, 13 (8.4%) had evidence of HSV acquisition prior to labor. Clinical manifestations defined two groups: 8 babies with early evidence of infection and 5 with late in utero acquisition. Of the 8 infants, all were premature. Manifestations included skin lesions/scars at birth (5/8), chorioretinitis (2/4), microphthalmia (3/7), microcephaly (5/8), and other CNS abnormalities on CT scan (6/8). Two infants died; the 6 surviving had severe neurologic sequelae. Four mothers had a history of primary genital HSV infection between 6-16 weeks. Signs of infection in the latter group were skin lesions at birth (5/5), hepatosplenomegaly (3/5), and other organ involvement (3/5). Two were prematures. One died from HSV pneumonia while the other 4 had no documented CNS sequelae. For this group, rupture of membranes >4 hours occurred in 1 case. Only 1 mother had a history of genital HSV infection (28 weeks) but 3 fathers of other babies reported genital lesions. Other etiologies for congenital infection were excluded. These findings indicate transmission of HSV can occur at any time during gestation. Although skin lesions are the hallmark of infection, regardless of time of acquisition, surviving newborns of early in utero infection are likely to have severe neurologic sequelae.

Protection from genital herpes simplex virus type 2 infection by vaccination with cloned type 1 glycoprotein D.

Guinea pigs were vaccinated with truncated herpes simplex virus type-1 (HSV-1) glycoprotein D produced in the genetically engineered mammalian cell line gD10.2. Vaccinated animals formed antibodies that neutralized both HSV-1 and herpes simplex virus type 2 (HSV-2) in an in vitro neutralization assay. Vaccinated animals were challenged with HSV-2 by intravaginal infection. Animals that received the immunogen in Freund's complete adjuvant were completely protected from the clinical manifestations of genital HSV-2 infection. Animals that received the immunogen incorporated in alum adjuvants were partly protected from clinical disease; the infections that did develop were significantly less severe than those that occurred in control animals injected with adjuvant alone. The results demonstrate that immunization with a purified viral protein can provide significant protection against primary genital infection by HSV-2 in guinea pigs.

Recurrences after first episodes of genital herpes in patients treated with topical acyclovir cream

The effect of topical acyclovir treatment of first episode genital herpes on the time to first recurrence in a group of 42 patients receiving either acyclovir or placebo was investigated. Topical acyclovir treatment had no effect on time to first recurrence in patients with either first episode HSV-1 or HSV-2 infections. There was no significant difference in the time to first recurrence in patients with either true primary or initial genital infections. However, the time to first recurrence in patients with first episode HSV-2 was significantly shorter than in patients with first episode HSV-1. Acyclovir treatment appeared to have no effect on the development of neutralising antibody in patients with either virus type.