Cellular immunity in vaccinia infection of mice. Anti-thymocyte serum effects on primary and secondary responsiveness.

Rabbit anti-mouse thymocyte serum suppressed host cell-mediated responsiveness to intravenously administered vaccinia virus, thereby augmenting the morbidity and mortality of this infection. It did not affect either humoral antibody or interferon production in response to vaccinia virus. No effects were noted on primary or secondary immunity to intracerebral virus inoculation.

Genital herpesvirus hominis type 2 infection: an experimental model in cebus monkeys.

Genital infection with herpesvirus hominis type 2 was established in ten female cebus monkeys. Clinical and laboratory findings in the cebus mimic closely those observed in humans, thus providing an experimental model which may be used in the study of the possible role of genital herpetic infection in cervical cancer and in perinatal and chronic neurological diseases.

Tumor-associated antigens in cervical cancer tissues and in sera from patients with cervical cancer or with head and neck cancer.

Hyperimmune New Zealand White rabbit sera prepared against partially purified tumor-associated antigens (TAA) of invasive cervical cancer tissues (CaCx's) were used to demonstrate TAA in CaCx's and circulating TAA (C-TAA) in sera from patients with cervical cancer or with head and neck cancer. Anti-CaCx serum adsorbed with pooled normal cervical tissue (NCx) antigen preparations and with lyophilized pooled normal human plasma gave precipitin in gel reactions with CaCx but not with NCx, which indicated the presence of TAA in CaCx. The adsorbed antisera reacted with sera from patients with cervical and head and neck cancers, which indicated the presence of C-TAA in such sera. False-positive reactions with control sera from normal humans or from patients with benign gynecologic diseases were much less frequently observed. Statistical analysis of data obtained by the testing of 96 coded serum samples from the National Cancer Institute-Mayo Clinic Serum Bank (Rochester, Minn.) revealed that the results were significant and that specificity was high but sensitivity of the assay was relatively low.

Epidemiological studies relating genital herpetic infection to cervical carcinoma.

PIP: Epidemiological studies relating genital herpetic infection to cervical carcinoma are reviewed. The high frequency of herpes simplex virus type 2 (HSV-2) antibodies in young women (21 years or younger) with cervical carcinoma in situ and in women with dysplasia or carcinoma in situ, matched for various sexual attributes to control women, provide support for a causal relation. However, various laboratory, histopathological, and statistical problems associated with all epidemiological studies do not yet permit a firm conclusion as to the etiological role to the genital virus in cervical carcinogenesis. With the use of herpes-related cancer antigens or purified HSV-2 type-specific antigens, and with the possible development of protective HSV-2 vaccines, the application of epidemiological approaches may be necessary to provide the most finite evidence of causality.^ieng

Relation of cytohistopathology of genital herpesvirus infection to cervical anaplasia.

PIP: Cytological changes resulting from herpes simplex virus (HSV) were detected in 673 (.26%) of 279,589 cervicovaginal smears from an indigent hospital population. Histopathological features of HSV infection were observed in 45 of 126 biopsies taken from the group of 673. Most of the lesions were multifocal and were located on the squamocolumnar junction; diagnostic cells were found at the base of normal squamous epithelium or at the surface of anaplastic epithelium. The cytological and histological changes and diagnostic procedures for HSV infection are given in detail; cytological detection is 2/3 as sensitive as viral isolation. The presence of anaplasia in about 2/3 of the 673 HSV cases either before or at the same time as the detection of HSV suggests that anaplastic tissue is more susceptible to HSV infection. However, serological evidence showing HSV antibodies before the onset of cervical anaplasia, the recurring nature of HSV, and epidemiological evidence showing the age of acquisition of HSV to be 5-30 years before diagnosis of cervical cancer are all compatible with the view that HSV causes cervical cancer.^ieng

Rosette formation and inhibition in cervical dysplasia and carcinoma in situ.

Total and early rosettes and rosette inhibition were measured in patients with cervical dysplasia and carcinoma in situ. Both total and early rosettes were significantly depressed in patients with carcinoma in situ; early rosettes were also significantly lower than were controls in women with severe dysplasia. Rosette inhibition titers were increased in most patients with moderate dysplasia and in all patients with either severe dysplasia or carcinoma in situ. Thus, the Rosette inhivition test may be useful in detecting, in a precancerous state, patients at risk for cancer.

Immunology of herpes simplex virus infection: relevance to herpes simplex virus vaccines and cervical cancer.

The immunology of herpes simplex infections has been reviewed, particularly in relation to potential herpes simplex virus (HSV) vaccines, and to the association between HSV and cervical cancer. Relevant data from humans, experimental animals, and in vitro systems implicate both specific immune mechanisms and nonspecific factors in the course of HSV infections. There appear to be complex interactions between the various populations of mononuclear cells, marcrophages, and lymphokines or other humoral factors. It is not yet possible, however, to pinpoint the crucial factors determining the different manifestations of the viral infection (primary infection, endogenous recurrence, or exogenous reinfection) in the various human hosts, although genetic factors may be important. While numerous animal models of HSV infection are available for the evaluation of HSV vaccines, models of HSV-induced cervical cancer require further exploration.

A preliminary report on investigation of oncogenic potential of herpes simplex virus type 2 in Cebus monkeys.

A total of 301 female and 133 male Cebus monkeys have been placed under study during a 3-year period. Females are inoculated intradermally into the cervix every 6 months; 225 receive virus and 76 receive control material. More than 50% of the animals were infected on primary inoculation, and a similar percentage was foundon the 1st reinoculation. Males are housed with females at 1:1 to 1:3 ratios. Eighteen % of the males exposed to virus-inoculated females have become infected. To date, a total of 55 pregnancies have produced 14 live births and 8 abortions. The remaining 33 animals are still pregnant. No neonatal herpes simplex virus type 2 infections have been identified. Cytological changes of mild (atypia) to moderate (dysplasia) anaplasia have persisted for 12 to 32 months in 13 herpes simplex virus type 2-infected females. These animals received their 1st inoculation 14 to 50 months ago. Persistent anaplasia has not been found in control animals.

Correlation of virus load and soluble L-selectin, a marker of immune activation, in pediatric HIV-1 infection.

OBJECTIVE: HIV infections in children are characterized by high viral load and, in some perinatally infected newborns, delayed appearance of viral markers. Both phenomena may be related to different levels of immune activation affecting viral replication. This study was designed to investigate the relationship between immune activation and viral replication in pediatric HIV infection, and the role of pre-existent immune activation in facilitating HIV transmission to the fetus/newborn. DESIGN: Plasma levels of soluble L-selectin (s-LS), an immune activation marker, were determined in 100 infants with perinatally transmitted HIV infection, compared with 106 age-matched HIV-exposed uninfected controls. Included in the analysis were samples from 31 HIV-infected (10 PCR+ and 21 PCR-) and 35 uninfected newborns aged < 2 days. METHODS: To determine s-LS levels, a solid phase ELISA was performed on plasma samples of patients and controls. RESULTS: s-LS levels in uninfected children were higher than those in normal adults. HIV-infected patients had more rapidly increasing values in the first 6 months of life compared with uninfected infants. Plasma s-LS levels correlated with HIV viral loads (r, 0.50). Among newborns in the first 2 days of life, s-LS levels were lowest in those with negative PCR tests, compared with PCR-positive or uninfected infants. CONCLUSIONS: These results suggest that higher immune activation in children contributes to higher viral loads, and that the level of pre-existent immune activation may have a role in determining which infants have detectable virus in peripheral blood at birth.

A prospective population-based study of HIV perinatal transmission.

OBJECTIVE: To estimate the perinatal HIV transmission rate and describe the natural history of infant HIV infection in a situation in which HIV status is known in more than 95% of delivering women. DESIGN: A cohort of HIV-exposed infants born between 7 July 1987 and 30 June 1990, whose mothers were identified by routine voluntary universal HIV testing, were followed using clinical and laboratory measures. SETTING: Grady Memorial Hospital, a major health-care site for individuals of lower socioeconomic status in Atlanta, Georgia, USA, with approximately 7000 deliveries per year. PATIENTS: HIV-exposed infants (n = 165), 98% of whom were African American. RESULTS: Annual maternal HIV seroprevalence increased from 0.58 to 0.86%. The annual proportion of HIV-positive women having a second delivery increased from 4.3 to 25%. Clinical outcome was known for 132 out of 165 infants (22 infected and 110 uninfected), the transmission rate was 17% (confidence interval, 11-24%). The rate declined to 11% by the third year of the study. Gestational growth, prematurity and mode of delivery were unrelated to infant outcome. There was a trend for intravenous drug use to be more common in mothers of infected infants (P = 0.08). After 35 months median follow-up of infected infants, eight out of 22 (36%) had an opportunistic infection (seven Pneumocystis carinii pneumonia); three out of 22 (14%) had lymphocytic interstitial pneumonia, and 10 out of 22 (45%) were asymptomatic or had only nonspecific symptoms. Cumulative mortality in infected infants was 9, 32 and 32% by 1, 2 and 3 years of age, respectively. CONCLUSION: In this cohort of HIV-exposed infants, perinatal HIV transmission was 17% overall. Factors affecting the transmission rate and possible future changes in the rate require further study.

A simple perfusion technique for isolation of maternal intervillous blood mononuclear cells from human placentae.

A noninvasive perfusion method for the recovery of maternal placental (intervillous) blood for use in immunologic assays is described. 60% of the perfused blood samples tested for fetal red blood cell (RBC) contamination were found to be pure maternal blood; in the remainder, fetal RBC contamination, with a single exception, was less than 6%. The intervillous mononuclear cells (IVBMC) isolated from this blood were of predominantly maternal origin as demonstrated by a polymerase chain reaction-based DNA typing technique. The number of IVBMC obtained was within the range of 9 to 55 X 10(6) cells. Phenotypic analysis of IVBMC surface antigens revealed that 61% of the cells were CD3 + T-cells and 18% were CD19 + B-cells. The CD4 + and CD8 + T-lymphocyte subsets accounted for 28 and 26% of the IVBMC, respectively. The IVBMC were functionally competent as evidenced by in vitro lymphoproliferation and cytokine production in response to mitogen and PPD stimulation. This technique allows for rapid and safe isolation of large numbers of IVBMC which are functionally active up to 12 h post-delivery, thus representing a significant improvement over previously described methods. It should facilitate more vigorous research in the study of uteroplacental immunity and infectious disease research, particularly in field settings where sample collection and laboratory facilities are distant.

Different in vitro effects of dual combinations of anti-herpes simplex virus compounds.

Five promising antivirals have been tested individually and in pairs on herpes simplex virus (HSV) types 1 (Strain F) and 2 (Strain G) in Vero cells. These are: 9-(2-hydroxyethoxymethyl)guanine (acyclovir, ACV), 9-beta-D-arabinofuranosyladenine (ara-A), 5-trifluorothymidine (TFT), E-5-(2-bromovinyl)-2'-deoxyuridine (BVDU), and phosphonoformate (PFA). Various types of interactions depending on virus type were observed: synergistic [ara-A/TFT; ara-A/BVDU (G); BVDU/PFA (F)]; additive [ACV/ara-A; ACV/TFT; ACV/BVDU; ACV/PFA (G); BVDU/TFT; PFA/ara-A; PFA/TFT]; and sub-additive [ACV/PFA (F); ara-A/BVDU (F) and BVDU/PFA (G)]. Neither antagonism nor interference was noted for any combinations.

Patient-initiated therapy for recurrent genital herpetic infections.

Therapy for recurrent genital herpes is difficult to evaluate because of the short duration of lesions and viral shedding. Very early treatment, however, can be begun by patients who experience prodromes before onset of lesions. We have found prodromes to occur in 73 percent of male and 84 percent of female patients; 57 percent of the men and 68 percent of the women experienced prodromes in at least three out of every four recurrent episodes. Variables that might affect results of such a patient-initiated study evaluating topical acyclovir included: application of the ointment (drug or placebo) without prodrome (10 percent); the possibility of a false prodrome (less than or equal to 10 percent); long intervals between experiencing a prodrome and ointment application (15 percent greater than 24 hours); the inability to sense a prodrome during sleep; lack of ointment application to the involved areas with initial or later lesions (approximately 15 percent); noncompliance of enrollees in returning for study evaluations (greater than or equal to 10 percent). Such variables will need to be considered when the code is broken in this multicenter study, as well as for patient-initiated studies with other formulations of acyclovir or with other drugs. Earlier negative studies with other agents given within two to three days of appearance of lesions might also require reevaluation with similar patient-initiated studies.

Rapid detection of viruses in biopsy or autopsy specimens by the pseudoreplica method of electron microscopy.

A rapid pseudoreplica method of electron microscopy (EM) was used to detect viruses in brain tissues obtained from 57 patients suspected of having herpes simplex encephalitis (HSE). Herpesvirus particles were demonstrated in 18 of the 20 (90%) specimens from which herpes simplex virus was isolated in tissue culture. The procedure takes less than 2 hours, as compared to the time (greater than or equal to 2 days) usually required for virus isolation or standard thin-section electron microscopy. Although the immunofluorescence test has comparable rapidity, it was found to be less sensitive and to yield occasionally false-positive reactions. Our findings indicate that, until a brain biopsy can be replaced by less invasive procedures, examination of brain tissue by the pseudoreplica method offers a rapid diagnosis of a herpesvirus encephalitis. Furthermore, the demonstration of viruses in brain, lung and liver specimens obtained by biopsy or at autopsy suggests that this electron-microscopic method would be advantageous for detecting viruses in tissue samples, particularly in the case of viruses which may not be readily demonstrable by conventional methods.

Antibody-dependent cellular cytotoxicity to target cells infected with herpes simplex viruses: functional adequacy in the neonate.

The functional adequacy of antibody-dependent cellular cytotoxicity (ADCC) in the human neonate was evaluated in a 51Cr release assay which employs tissue culture cells acutely infected with type 1 or type 2 herpes simplex virus (HSV) as targets. Two aspects of ADCC were assessed: cytotoxic effector activity in cord blood mononuclear cells (MC) and the ability of the antibody mediating ADCC to pass the placenta. Although effector cell activity was readily detected in all 13 cord blood specimens tested, cord blood MC showed moderately reduced cytotoxic activity when compared with blood MC from normal adults at the same effector cell:target cell ratio. This finding suggests that effector cells in cord blood make up a reduced proportion of the total circulating MC population and may be of relevance to the newborn's increased susceptibility to HSV infection. On the other hand, the number of MC in cord blood was found to be almost twice that of adult blood, suggesting that the absolute number of ADCC effector cells in cord blood was within the adult range. The antibody mediating ADCC to HSV-infected cells was shown to be transferred quantitatively across the placenta, providing further evidence that it is an IgG immunoglobulin.

The natural history of herpes simplex virus infection of mother and newborn.

The controlled evaluation of vidarabine as therapy of neonatal herpes implex virus (HSV) infection provided an opportunity to collect data to further assess the natural history of maternal and newborn infections. Women delivering infected babies were young, nulliprous, and infrequent aborters. Nearly 50% of the gestations ended in premature labor. Maternal infection was asymptomatic in 39 of 56 (70%) of the mothers, at the time of delivery. However, risk factors included a past history of genital herpes at any time and exposure to a sexual partner with presumed HSV lesions. Associated diseases in children born to these women were common. Premature infants had an incidence of respiratory distress of 52% (14 of 27). Eight of 29 (28%) term newborns had a bacterial infection, antedating the onset of neonatal HSV infection. Virologic studies on infected newborns confirmed that skin lesions were the most frequent site for virus retrieval. Progression of disease from isolated skin lesions was common, occurring in 70% of babies whose presenting sign was skin vesicles. CSF was virus-positive from 14 babies and more frequently in those with localized CNS disease. Importantly, brain biopsy was necessary for diagnosis in four cases. Finally, neither the presence or absence of antibodies to HSV was useful in predicting either presentation or outcome of infection. These studies further emphasize the complex nature of HSV infections of the newborn and need for tertiary care.

Neonatal herpes simplex virus infection: follow-up evaluation of vidarabine therapy.

An open study of vidarabine (adenine arabinoside) therapy was performed to verify the mortality from neonatal herpes simplex virus infection and to define further long-term morbidity. A total of 39 babies not previously reported were treated with either 15 mg/kg/d (16 newborns) or 30 mg/kg/d (23 newborns) of vidarabine administered intravenously for ten to 14 days. Outcome was compared with that from 56 newborns evaluated in a prior trial. Irrespective of the dose of medication, therapy decreased the mortality in babies with disseminated and CNS disease to 40%. The extent of organ involvement and, in particular, pulmonary herpes simplex infection were predictive of mortality (P = .001, for both). For these babies, 32% achieved normal developmental milestones 2 years after therapy. Disease localized to the skin, eye, and/or mouth was not associated with death. However, neurologic impairment occurred in 12% of this treated group of newborns. These findings underscore the value of vidarabine therapy of neonatal herpes simplex virus infection. However, an increase in dosage did not appear to result in significant improvement in either mortality or morbidity. Further improvement in the mode of therapy and the utilization of more potent antiviral drugs are currently being tested.

Vidarabine therapy of neonatal herpes simplex virus infection.

Vidarabine (adenine arabinoside) was evaluated for treatment of neonatal herpes simplex virus infection in a randomized controlled study. Of 56 infected newborns, 13 had infection of skin, eye, or mouth only, 16 had localized brain disease (CNS), and 27 had disseminated disease. Both treatment and placebo groups were comparable by disease distribution and for major population characteristics. Because of the severity of CNS and disseminated disease, these groups were combined for mortality assessment. Mortality was significantly reduced in babies with CNS and disseminated disease from 74% to 38% with drug therapy, P = .014. Outcome in babies with disseminated disease alone, although improved, was poor. Death rate was reduced from 85% to 57% with therapy. Only 14% of drug and 8% of placebo recipients were assessed as normal at 1 year of age. Outcome was better with localized CNS disease; mortality was reduced from 50% to 10%. With treatment, 50% of infected newborns were normal and without only 17%. With skin, eye, or mouth infection death did not occur; however, severe sequelae occurred in 38% of placebo and minor sequelae in 25% of drug recipients. No evidence of acute toxicity was identified in this study. Thus, a beneficial effect of vidarabine therapy on neonatal herpes simplex infection is similar to that evident with therapy of herpes simplex encephalitis occurring in older individuals. Nevertheless, improvement in the mode of therapy or the development of more potent antiviral drugs is essential.

Comparative study of diagnostic procedures for congenital cytomegalovirus infection.

In a prospective study the incidence of congenital cytomegalovirus (CMV) infection was 2.2% (31 OF 1,412) as evidenced by viruria during the first week of life. Among immunoserologic methods used to screen these neonates, the rheumatoid factor test, although non-specific, proved to be the most convenient; its sensitivity for identifying infants with CMV infection was 35% to 45% with no false-positives. The rates for correct and incorrect identification of neonates at risk was, respectively, 33% and 3.1% when testing for increased levels of IgM; 5% and 10% when testing for increased levels of IgA; 76% and 21% when testing for IgM anti-CMV (IgM immunofluorescent test) antibody, and 0% when testing for IgA anti-CMV antibody. Rapid virologic diagnosis was achieved by assessing urine specimens. Confirmation by electron microscopy was possible in less than one hour in 92% of cases. The detection of early induced CMV-specific nuclear antigens by anticomplement immunofluorescence was diagnostic in 91% of cases within one day of inoculation of specimens in tissue culture. Infectivity of CMV in urine was well preserved for a least seven days at 4 C. Thus, in order to achieve a rapid diagnosis of congenital CMV infection, in sick as well as asymptomatic neonates, urine specimens may, if necessary, be transported at 4 C to distant laboratories.

Antiviral chemotherapy and neonatal herpes simplex virus infecition: a pilot study--experience with adenine arabinoside (ARA-A).

Among 13 neonates with herpes simplex virus (HSV) infection, eight had disseminated disease, one localized CNS disease, and in four the infection was confined to the skin and eyes. Ara-A, a purine nucleoside with anti-viral activity against DNA-VIRUSES, WAS GIVEN (10 TO 20 MG/MG/DAY) BY A CONTINUOUS 12-HOUR INTRAVENOUS DRIP FOR 10 TO 15 DAYS. In all, ara-A administration was begun within three to eight days after the appearance of skin vesicles which represented the hallmark of the disease. Both diagnosis and ara-A treatment were much delayed in one infant without skin vesicles and four infants whose skin vesicles appeared late, long after the occurrence of CNS damage. In this group of infants with disseminated disease, four died and one infant was left with severe neurological deficits. Eight infants (four with disseminated and four with localized skin disease) with skin vesicles as the earliest sign of infection received ara-A early, within three days after the onset of neurologic signs. All survived with no neurologic deficit at 6 months to 1 year of age. There was no apparent toxicity of ara-A to the bonemarrow, liver, or kidney.