Israel Sarov

Placebo-controlled trial of topical interferon in labial and genital herpes

The efficacy of topical interferon-beta (IFN-beta) treatment was assessed in 25 patients with herpes of the lips or genitals who completed a 2-year follow-up in a double-blind placebo-controlled trial. IFN-beta gel (10(5) U/g) 4 times daily (about 2 x 10(4) U) applied locally during eruptions (about 10 days) reduced the mean number of recurrences (p less than 0.007) and the duration of eruptions (p less than 0.007): in the placebo group these indices did not change significantly. Reduction of symptoms and severity was noted in 11 of 12 patients on IFN-beta and in only 1 on placebo. No important side-effects were recorded. Topical IFN-beta may therefore be advantageous as a time-limited local treatment of recurrent herpes simplex virus infections of the genitals and lips.

The association between chlamydial-specific IgG and IgA antibodies and pregnancy outcome in an in vitro fertilization program

Chammydial-specfic IgG and IgA antibodies were determined by a single serovar (L2) immunoperoxidase assay (IPA) in the serum of all patients that have conceived in an in vitro fertilization and embryo transfer (IVF & ET) progrom (n=106) and in a group of patients that went through the program at the same period of time and did not conceive (n=94). The prevalence rate of elevated IPA IgG (titers≥1∶128) and IPA IgA (titers≥1∶16) specific to chlamydiae was significantly higher (P<0.001) in the IVF&ET pregnancy loss and nonconception groups (“failures”) versus the IVF&ET term pregnancy group (“successes”) (74 vs 47%, odds ratio=4.1, and 34 vs 14%, odds ratio=4.3, respectively). Stepwise discriminant analysis revealed that elevated specific chlamydial IgG had the greatest effect on the variance between successes and failures in this study group. Our study indicates the possible role of past or chronic active chlamydiae infection on the “take-home baby rate” in an IVF&ET program.

A rapid immunoperoxidase assay for the detection of specific IgG antibodies to Chlamydia trachomatis.

A technique, using indirect immunoperoxidase antibody (IPA), was developed for the detection of IgG antibody to Chlamydia trachomatis. The IPA technique employs glass slides with air-dried and acetone-fixed C trachomatis infected cells, which can be stored at -70 degrees C and used for several months. Antibody titres detected by IPA were comparable to those detected by the indirect fluorescent antibody technique.

Detection of Specific IgA Antibodies in Serum of Patients with Varicella and Zoster Infections

A sensitive solid-phase, enzyme-linked immunosorbent assay (ELISA) was developed for detection of serum IgA antibodies to varicella-zoster virus (VZV). The antigen consisted of a sonically disrupted extract of VZV-infected human embryo cells. Rabbit antihuman IgA peroxidase conjugate was used to detect human IgA bound to viral antigens. In parallel, IgM and IgG antibodies to VZV were studied by ELISA and by an immunoperoxidase antibody to membrane antigen technique, respectively. VZV IgA antibodies were detected in high titers in all 5 varicella and 8 zoster patients. Specific VZV IgM antibodies were detected in all 5 varicella patients, but only in 2 of 8 zoster patients. No VZV IgA antibodies (less than 40) were detected in 50 healthy control sera. Neither were they found in paired sera of 5 patients with herpes simplex infections, 2 patients with Epstein-Barr virus infections, and 5 patients with human cytomegalovirus infections. The potential application of ELISA IgA techniques in serodiagnosis of both primary and reactivated VZV infections is discussed.

Tryptophan reversal of recombinant human gamma-interferon inhibition ofChlamydia trachomatis growth

Recombinant human gamma-interferon was shown to inhibit the growth ofChlamydia trachomatis (L2/434/Bu) in HEp-2 cells. This inhibition could be reversed by the addition of tryptophan. The effect of tryptophan was dose dependent and determined by the interferon concentration. At low concentrations of interferon, the addition of tryptophan completely restoredC. trachomatis infectivity, whereas at high concentrations (100–1000 IU/ml) the effect of interferon could not be totally reversed. The reversal effect of tryptophan could be achieved even when the addition was 48 h after infection. The probability that tryptophan degradation induced by gamma-interferon might be the mechanism involved in its antichlamydial activity is discussed.

Isolation and polypeptide characterization of varicella-zoster virus

Abstract Varicella-zoster virions (VZV) were isolated from cytoplasm of VZV-infected human fibroblasts by zone centrifugation in glycerol gradients followed by two cycles of equilibrium flotation on glycerol-tartrate gradients. The purified particles were characterized with respect to their morphology and polypeptide composition. VZV was found to be composed of at least 33 species of polypeptides which were identified by means of SDS-polyacrylamide gel electrophoresis and autoradiography. They ranged in molecular weights from 16,000 to 244,000. Glycosylated components were identified by SDS-gel electrophoresis and autoradiography of purified [ 14 C]glucosamine-labeled virus. VZV virions were found to contain at least 5 glycopeptides ranging in molecular weights from 140,000 to 52,000.

Detection of IgG and IgA antibodies to Chlamydia trachomatis in sera of patients with chlamydial infections: use of immunoblotting and immunoperoxidase assays

The immune response to individual structural polypeptides of Chlamydia trachomatis was studied in 75 sera from symptomatic and asymptomatic women with culture-proved genital infections and from apparently healthy women who were culture-negative for C. trachomatis. The immunoblotting technique and the single serovar (L2) inclusion immunoperoxidase assays were used for measurement of the various antibodies. Antibodies to 18 structural polypeptides, ranging in molecular weight from 29 to 204 Kdaltons, were detected by the immunoblotting technique in sera from seropositive women. The immunoperoxidase assay showed that sera with high titers of IgG and IgA antibodies to C. trachomatis reacted with more polypeptides than did sera with low titers in this test. Antibodies to the 60- and 62-Kdalton polypeptides were detected in almost all sera positive for IgG and IgA antibodies, irrespective of chlamydial shedding. About 40% of sera with high IgG and IgA titers reacted with 39-, 57-, 64-, 72-, 86-, 105-, 155-, and 204-Kdalton polypeptides. The prevalence of IgA antibodies to C. trachomatis was higher among women with culture-proved chlamydial infections than among apparently healthy controls.

Human cytomegalovirus specific IgA antibodies detected by immunoperoxidase assay in serum of patients with cytomegalovirus infections

Cytomegalovirus (CMV)-specific IgA antibodies were determined by an immunoperoxidase assay in sequential serum samples of 10 patients with CMV infection in order to evaluate the feasibility of the use of this technique for diagnosis. In parallel, IgM and IgG antibodies to CMV were studied by enzyme-linked immunosorbent assay (ELISA) and by the immunoperoxidase assay, respectively. CMV IgA antibodies were detected in all 10 CMV patients studied. Specific IgM was detected earlier than IgA in only one of these ten patients. No CMV-specific IgA antibodies (titer less than 2) were detected in 45 medical students. Neither were they found in paired sera of 5 patients with herpes simplex infection, 5 patients with varicella, 6 patients with zoster and 2 patients with Epstein-Barr virus infection. The potential application of the indirect immunoperoxidase IgA assay for serodiagnosis of CMV infections is discussed.

Detection of IgG Antibodies Specific for Measles Virus by Enzyme-Linked Immunosorbent Assay (ELISA)

A solid-phase, enzyme-linked immunosorbent assay (ELISA) for determination of IgG antibodies against measles virus is described. The assay utilized antigen-coated polystyrene microplates. The antigen consisted of a sonicated extract of measles-infected Vero cells. Goat and anti-human IgG-peroxidase conjugate was used to detect human IgG bound to viral antigen. Sera taken from 63 healthy adults, 11 young children and 36 patients were evaluated for their IgG titer against measles virus. Comparison of results obtained by ELISA with those obtained by hemagglutination-inhibition (HI) assay or by complement fixation showed good agreement between the tests. The geometric mean titer (GMT) for healthy adults was 753 for ELISA and 32.8 for HI. If these averages are taken as a measure of comparison, then ELISA is approximately 23 times more sensitive than HI. ELISA technique is rapid to perform and could be recommended for routine diagnosis.

Persistence of antichlamydial antibodies after treatment of acute salpingitis with doxycycline

The effect of treatment with doxycycline on serum IgG and IgA antichlamydial antibodies was evaluated in 33 women who had had acute salpingitis associated with high titers of serum IgG (> or = 1:128) and/or IgA (> or = 1:16) antichlamydial antibodies. Overall, 29 women (87.9%) remained with high titers of IgG and/or IgA antibodies. No change or insignificant change in IgG antibody titer was demonstrated in 21 women (63.6%) and in IgA antibody titer in 21 women (63.6%). Positive seroconversion or a significant increase (> or = 4-fold) in IgG antibody titer was demonstrated in eight women (24.2%) and in IgA antibody titer in six women (18.1%). Negative seroconversion or a significant decrease in IgG antibody titer was demonstrated in four women (12.1%) and in IgA antibody titer in six women (18.1%). It is concluded that in most patients who had acute salpingitis associated with pretreatment high titers of serum antichlamydial antibodies, posttreatment titers may remain high even if treatment with doxycycline results in complete resolution of clinical signs and symptoms of the disease.