Cross-sectional study of immunologic abnormalities in intravenous drug abusers on methadone maintenance in New York City.

One hundred and ninety-nine patients with a history of intravenous drug abuse, and enrolled on the St Luke's-Roosevelt Hospital Center Methadone Program, had baseline evaluations performed from September 1984 to April 1987. The study was designed to examine immunologic parameters associated with HIV seropositivity and those predictive of progression to AIDS-related complex (ARC) and AIDS. Sixty-four patients (32%) had antibodies to HIV by enzyme-linked immunosorbent assay (ELISA), with confirmation by Western blot and none of these patients had ARC or AIDS at the time of initial evaluation. The mean values for white blood-cell count, absolute lymphocyte count, proportion and absolute CD4, and CD4/CD8 ratio were decreased significantly in the HIV-seropositive group compared with the HIV-seronegative group. On the other hand, levels of circulating beta 2-microglobulin, SCD8, SIL-2R, and HIV p24 antigen were significantly elevated in the HIV-seropositive group compared with the HIV-seronegative group.

Impaired cell-mediated immune response in patients with congenital rubella: correlation with gestational age at time of infection.

Lymphocyte transformation, interferon, and leukocyte migration inhibition factor synthesis were studied in purified lymphocyte cultures for 20 children with congenital rubella and 18 healthy children (seven susceptible and 11 immune to rubella). Lymphocyte transformation after phytophemagglutinin stimulation was significantly lower in children with congenital rubella as compared to healthy controls. Responses to purified rubella virus were absent in the susceptible controls and absent or at least two times lower in congenital rubella children than in immune controls. After purified rubella virus stimulation, leukocyte migration inhibition factor production was detected in all immune controls, but in none of the susceptible controls, or the congenital rubella-infected children. The results varied with gestational age of intrauterine infection: the impairment of cellular immune response, both after phytohemagglutinin or rubella virus stimulation, was more severe in the children infected in the first two months than in the latter stages of gestation.

Inhibition of HIV replication in lymphocyte cultures of virus-positive subjects in the presence of sho-saiko-to, an oriental plant extract.

An oriental remedy, Sho-saiko-to (SST) consisting of a mixture of aqueous extracts from seven different plants and whose most active component is the chemically defined compound baicalein was tested for its ability to inhibit the production of the human immunodeficiency virus (HIV). The testing was done with cultures of human lymphocytes obtained from HIV-positive asymptomatic subjects and patients with ARC or AIDS. The replication of the virus was monitored by quantitative assay of the reverse transcriptase (RT) activity and of the synthesis of antigen p24. The lymphocyte cultures (LC) were maintained in the absence and in the presence of 25, 50 or 100 micrograms/ml of SST, and monitored for up to 5 weeks. The results showed that in LC from asymptomatic subjects RT activity and synthesis of p24 was completely inhibited by low concentrations of SST. High concentrations of SST inhibited virus replication in 80% of LC from ARC patients, but were completely ineffective in LC from AIDS patients. It was observed that the RT activity was more sensitive to inhibition by SST than the synthesis of p24, and that the antiviral effect was dependent on the virus load of the LC.

Open study of AL-721 treatment of HIV-infected subjects with generalized lymphadenopathy syndrome: an eight week open trial and follow-up.

AL-721 is a lipid compound composed of neutral lipids, phosphatidylcholine and phosphatidylethanolamine in a 7:2:1 ratio. The objective of this open study was to evaluate the effects of AL-721 in vivo in an 8-week open trial in which 10 g twice daily was administered on a low fat diet to eight HIV-infected subjects with lymphadenopathy syndrome (LAS). Serial lymphocyte cocultivation studies in 7 patients with initial culture positivity appeared to demonstrate reduction of reverse transcriptase peak counts in 5 with the trough noted in 4 at 8 weeks and in one at 4 weeks following termination of therapy. The mean values for all 7 patients revealed a baseline value of 73,419 with decrease to a low of 27418 at 8 weeks. Mean levels of total lymphocytes, T-4, T-8 and T-11 cells were not altered but lymphoproliferative responses to concanavalin A and pokeweed mitogens appeared to be augmented in 4 of the 8 subjects in association with AL-721 treatment. No side effects were noted. In a subsequent follow-up study using a normal diet in the same subjects lymphocyte cocultivation and mitogen-induced responses were less consistently affected when 15 g twice daily AL-721 was readministered. In addition, serum HIV p24 antigen and CD4 levels were not altered during both the 8-week open and subsequent AL-721 readministration. Four of the 8 patients have progressed to AIDS over the subsequent 14 months.

Prospective observations of viral and immunologic abnormalities in homosexual males.

PIP: This article presents preliminary observations on the immunologic and viral status of 103 homosexual men from New York who were enrolled in a prospective study begun in late 1981. At the time of enrollment, none of the study participants had acquired immunodeficiency syndrome (AIDS) as defined by the Centers for Disease Control (CDC). The initial laboratory investigation indicated that 82.7% of the 1st 81 volunteers had a OKT4:OKT8 ratio lower than the control range of 1.4-2.8. 31 (38.3%) of these 81 volunteers had 1 or more of the following signs and symptoms: history of fever, weight loss, diarrhea, lymphadenopathy, recurrent amebiasis, or persistent amebiasis. As part of the initial study, intensive laboratory investigations were performed on 30 of these subjects, including 20 who were symptomatic. Findings included abnormal T-cell markers, elevated immunoglobulins, circulating immune complexes, and cryoglobulins, as well as B-cell dysfunction. Follow-up of these patients 1 year later indicated that there was no significant improvement in T4:T8 ratio, and those volunteers originally identified as symptomatic continued to contribute the most abnormal ratios. A decrease in the absolute number of T4 lymphocytes contributed most to the ratio reversal. In addition, 26 of the 30 volunteers underwent viral isolation studies from 5 body sites or secretions. 23 of 26 were positive for virus from 1 or more sites compared with 3 of 33 heterosexual controls. Isolation of cytomegalovirus, especially from semen, was most frequent. It remains unclear whether the abnormal immunologic findings in healthy homosexual volunteers represent a transient response to viral infection, an epiphenonomenon due to multifactorial causes that make subjects susceptible to an unidentified AIDS agent, or AIDS itself. The cohort study will continue.^ieng

Immunosuppression and isolation of rubella virus from human lymphocytes after vaccination with two rubella vaccines.

Two groups of young rubella-susceptible women were vaccinated with two rubella vaccines. Heparinized blood samples were taken from all individuals the day of vaccination and 5, 7, 15, 21, 30, 35, and 42 days later. Purified lymphocytes from these samples were cocultivated with AGMK cells for rubella virus isolation. Parallel samples of lymphocytes were stimulated with phytohemagglutinin, and the rate of [14C]thymidine incorporation was determined. Rubella virus was isolated from lymphocytes collected on days 7, 15, and 21 after RA27/3 vaccination in contrast to days 7 to 35 after HPV77 vaccination. The lymphocyte response to phytohemagglutinin was markedly suppressed from day 5 to 15. Normal lymphocyte responses were restored within 1 month after vaccination with RA27/3, but even later (1 week) after HPV77 vaccine. Lymphocytes from rubella-susceptible persons infected invitro with rubella virus vaccines and stimulated with phytohemagglutin displayed a decrease in their responsiveness to the mitogen similar to that observed with lymphocytes from vaccinees. The transient immunosuppression observed in vaccinees is probably due to virus-induced functional damage of the lymphocytes since no direct cytocidal effect of rubella vaccine has been demonstrated on human lymphocytes.

Lymphocyte responses to rubella antigen and phytohemagglutinin after administration of the RA 27/3 strain of live attenuated rubella vaccine.

Lymphocyte phytohemagglutinin (PHA) responsiveness was found suppressed in both rubella sero-negative and sero-positive recipients of RA 27/3 strain of live attenuated rubella vaccine; the suppression was readily demonstrable only when a suboptimal dose of PHA was applied in the test. Lymphocytes from sero-negative vaccinees, which initially showed little or no in vitro response to concentrated rubella virus, became responsive after vaccination by day 21, when the highest sensitization to rubella antigen was seen. In the sero-positive vaccinees. lymphocytes responded to rubella antigen in vitro before vaccination, and in most cases vaccination did not result in significant changes in lymphocyte response. These results suggest that rubella vaccination leads to temporarily increased lymphocyte reactivity to rubella antigen, and the increased lymphocyte response to specific antigen may occur at the time of mild suppression of PHA response.

Cell-mediated immune response in rubella infections.

Cultures of lymphocytes from rubella-susceptible persons did not respond to stimulation with purified rubella antigen whereas those from persons who had experienced natural infection responded strongly to specific antigenic stimulation. The response of cultures of lymphocytes from vaccinees is weaker than that of naturally infected persons, and it appears that the response of lymphocytes from congenitally infected children depends on their gestational age at the time of interuterine infection. Vaccination or revaccination of immune subjects elicits a fast booster-type cell-mediated immune response. This property is useful for detecting the immune status of women exposed to rubella during pregnancy whose rubella history is unknown and whose humoral immune response is negative.

Adenosine in the treatment of recurrent herpes labialis.

The effectiveness of adenosine 5'-monophosphate was tested in thirty-six patients with recurrent herpes labialis. The compound displayed an immediate effect upon the healing of the recurrent lesions. Furthermore, twenty-three patients were free of recurrences for varying intervals extending over 2 years. In the remainder of the treated patients, a single recurrent episode was observed.

Interferon production in lymphocyte cultures after rubella infection in humans.

When stimulated with rubella virus irradiated with ultraviolet light, cultures of lymphocytes from individuals immune to rubella responded with production of 12-14 times more interferon than did cultures of lymphocytes from individuals susceptible to rubella. This property may be retained throughout life, since it was seen in individuals who had had rubella in their recent and remote pasts. Peak levels of interferon were detected six days after stimulation of lymphocytes with rubella virus antigen. In contrast, phytohemagglutinin induced production of interferon equally well in cultures of lymphocytes from persons immune and persons susceptible to rubella, with levels peaking three days after stimulation. Since there was a positive correlation among titers of hemagglutination-inhibiting antibody to rubella virus, incorporation of [14C]thymidine, and production of interferon after stimulation of lymphocyte cultures by rubella virus, interferon synthesis may be considered to be another measurable parameter of cell-mediated immunity in rubella.

Low level rubella immunity detected by ELISA and specific lymphocyte transformation.

The level of humoral and cell mediated immunity in persons with low or undetectable hemagglutination-inhibition (HAI) rubella titers was investigated by ELISA, IgG presence in sucrose gradient separated serum fractions and lymphocyte transformation. The study population consisted of persons with stated history of natural rubella infection and rubella vaccinees. Persons with natural rubella infection and HAI titers of 1:8 or +/- 1:8 (i.e., incomplete HAI at serum dilution of 1:8) were all ELISA positive and the stimulation index (SI) of specific lymphocyte transformation was higher than 2.5. Among the 20 persons with HAI titers of < 1:8, 8 were found to be ELISA positive and their SI was also > 2 and IgG was detected in their serum. Rubella vaccinees with HAI titers of 1:8 or +/- 1:8 were likewise ELISA positive. Their SI was lower: none higher than 3, but none lower than 1.5. Among 23 HAI negative vaccinees, 14 were found ELISA positive. This serum fraction contained IgG and the SI was > 1.5. It appears that ELISA test is able to detect antibodies where the HAI test fails. The positive outcome of ELISA test in this case was confirmed by the presence of IgG in serum fractions and by the lymphocyte response to rubella specific stimulation.

Efficacy of a virion envelope herpes simplex virus vaccine against experimental skin infections in hairless mice.

Hairless mice were immunized with herpes simplex virus type 1 (HSV-1) envelope antigen (EAG), EAG in association with polyriboinosinic . polyribocytidylic acid-poly-L-lysine complexed with carboxymethylcellulose (PICLC), and inactivated purified HSV-1 (VAG). After 2 weeks the mice were challenged by a percutaneous HSV-1 infection in the orofacial (OF) or lumbosacral (LS) skin area. Following immunization a consistent cell-mediated immune response was observed in all immunized mice, although the humoral immune response was very low, or not detectable. After challenge, a marked secondary humoral and cell-mediated immune response developed in all immunized mice, and the animals were protected against the development of skin lesions and the fatal outcome of infection. However, the establishment of latent infections in the sensory ganglia was not prevented by the immunization procedure.

Long-term follow-up of serum-interferon and its acid-stability in a group of homosexual men.

For a period of over two years 99 volunteer healthy homosexual men were examined periodically for the presence of interferon (IFN) in their serum. Thirty-nine subjects had either undetectable IFN levels in serum or IFN was detected only once in three to five samples tested. In another 45 subjects low IFN levels were detected throughout the study period. None of these subjects had or developed any disease symptoms. In the remaining 15 subjects high serum IFN levels were detected at their enrollment or during the study period. All these subjects started to manifest clinical symptoms compatible with AIDS. In six subjects the mean time elapsed between the first detection of serum IFN and disease symptoms was 6.5 months. In all subjects but one, the IFN was of type alpha. The acid-stability of serum IFN alpha decreased with time, and when its decrease was abrupt it was associated with a more rapid evolution of AIDS. Sera containing acid-labile IFN alpha can induce IFN alpha synthesis in normal lymphocyte cultures (LC), but do not influence IFN gamma synthesis in LC stimulated with phytohemagglutinin. LC stimulated with viral antigens in the presence of serum with acid-labile IFN alpha synthesized IFN with an increased sensitivity for acid treatment. The results confirm the prognostic value of serum IFN alpha in the development of AIDS, and suggest that the transition to acid-lability may be a gradual process.

Cell-mediated immunity in rubella assayed by cytotoxicity of supernatants from rubella virus-stimulated human lymphocyte cultures.

Rubella virus-stimulated lymphocytes from rubella-seropositive donors produced in the culture medium cytotoxic activity with preferential action against rubella-infected over uninfected target cells. The ability of lymphocytes to produce the cytotoxic activity upon stimulation by rubella virus correlated with the humoral rubella-immunity status, i.e. no such cytotoxic activity developed in the supernatants of lymphocyte cultures of rubella-seronegative donors. Stimulation of lymphocytes from seropositive donors by rubella virus was also detected by thymidine incorporation, but the correlation of lymphocyte responsiveness to the humoral rubella antibody status was not so clear as in the cytotoxicity assay. Conversion of lymphocytes from unresponsive to responsive to rubella virus following natural rubella infection and after rubella vaccination was demonstrated using both methods. Following vaccination rubella-specific cell-mediated immunity first became demonstrable at 14 days. The responsiveness of lymphocytes to phytohaemagglutinin (PHA) after rubella vaccination was followed by studying thymidine uptake and the ability of lymphocytes to produce lymphootoxin. By both tests marked suppression of PHA response occurred at days 3 and 7 after vaccination.

IgM detection by ELISA in the diagnosis of cytomegalovirus infections in homosexual and heterosexual immunosuppressed patients.

A new commercially developed cytomegalovirus (CMV)-IgM ELISA was found to be sensitive and specific when compared with sucrose gradient fractionation of Ig classes in CMV antibody-positive and negative sera. The presence of CMV IgM in patients' sera correlated with positive virus isolation from circulating mononuclear blood cells and urine. Serial examinations of patients with primary or recent CMV infection revealed a typical sequence of IgM and IgG development. The frequency of CMV isolation declined as the concentration of IgM decreased and the IgG levels increased. Since the isolation of CMV from clinical specimens is a cumbersome procedure, we suggest that the IgM ELISA could provide rapid and valuable information on the presence of an active or reactivated CMV infection.

Rubella-specific immune complexes after congenital infection and vaccination.

Circulating immune complexes which contained rubella-specific immunoglobulins were detected in 21 out of 63 subjects with congenital rubella and in 39 out of 65 subjects vaccinated with attenuated rubella virus, but in none of 43 subjects susceptible to rubella or 87 subjects with remote naturally acquired immunity to rubella. The presence or level of circulating immune complexes and the presence of rubella-specific complexes did not correlate with conventional serum rubella hemagglutination inhibition antibody titers. In the group with congenital infection, the presence of specific complexes many years after birth was associated with late-emerging clinical problems involving several organ systems. In vaccinates, the presence of specific complexes was associated with a higher incidence of side reactions. Two-thirds of the vaccinates and all of those revaccinated showed specific immune complexes as late as 8 months after immunization.

Virus isolation and immune studies in a cohort of homosexual men.

Virus shedding was detected in 77% of homosexual subjects and in only 6% of heterosexual controls. The overall virus isolation rate in homosexual subjects was not significantly different among HIV-seropositive (79%) and HIV-seronegative (74%) individuals. In about 20% of homosexual subjects, virus shedding from multiple sites was observed. The most frequently isolated virus was cytomegalovirus (CMV) (41%), followed by enteroviruses (23%), herpes simplex virus (HSV) (7%), and adenoviruses (6%). In the control group, about 50% of subjects were seronegative for HSV-1 and 2, and about 70% were negative for CMV and Epstein-Barr virus (EBV). Only 2% of homosexuals were seronegative for CMV, about 5% for HSV-1 and 2, and about 20% for EBV. No differences were found in antibody levels against varicella-zoster virus (VZV) among the control and homosexual groups. The proportion of seronegatives for Coxsackie and hepatitis viruses was significantly higher in control than in homosexual subjects. However, no differences in the proportion of seronegatives for measles, mumps, and rubella were observed. No HIV-antibody-negative individual was detected with an OKT4/OKT8 ratio of less than 0.75. On the other hand, only HIV-positive subjects, with a ratio of less than 0.75, had high serum IFN alpha titers. The results suggest that the high rate of virus shedding among HIV-negative homosexual subjects might be a factor in the development of AIDS in this high-risk population.

Immune response and latent infection after topical treatment of herpes simplex virus infection in hairless mice.

Treatment of herpes simplex virus (HSV)-infected hairless mice with a 2% phosphonoacetic acid (PAA) ointment prevented the appearance of virus-induced skin lesions and subsequent central nervous system (CNS) involvement. Treatment started 24 h after infection significantly reduced the intensity of the skin lesions and also prevented CNS involvement. After four to six applications of PAA ointment, a moderate skin erythemia developed, followed by scaling and complete healing 7 days after cessation of the treatment. Mice treated early after HSV infection had low or undetectable levels of virus-specific antibodies but were completely resistant to reinfection. Early treatment prevented the development of a latent ganglionic infection, but treatment initiated 24 h after infection could not prevent the establishment of the latent infection. PAA-treated and HSV-infected mice with nondetectable levels of antibodies did not develop, with a single exception, a latent ganglionic infection unpon reinfection. The cell-mediated immune response determined by levels of [14C]thymidine incorporation in Ficoll-Hypaque-purified spleen lymphocytes cultures was low in PAA-treated mice; it increased slightly after challenge infection but was strong in mice that proved to harbor a latent HSV infection in the ganglia.

Study of the lymphocyte in vitro response to rubella antigen and phytohemagglutinin by a whole blood method.

A whole blood culture method was used to study lymphocyte in vitro responses to rubella antigen and to phytohemagglutinin (PHA) in rubella infection. The acute phase of infection in four cases was characterized by high spontaneous incorporation of 14C-thymidine in the cultures, unresponsiveness of lymphocytes to rubella antigen, and absence of response, or relatively low response, to PHA. Cells showing vigorous in vitro response to rubella antigen appeared at about two weeks after the onset of rash. Lymphocyte PHA response returned to normal by day 31. Three rubella vaccinees exhibited a similar response. The use of whole blood lymphocyte cultures stimulated with multiple doses of mitogen and with antigen appears to be a promising technique for studies of "general" and "specific" cell-mediated immunity in viral infections.

Multicenter evaluation of a 1-h enzyme-linked immunosorbent assay for rubella serology.

A 1-h enzyme-linked immunosorbent assay (Rubestat) was developed for rubella virus immunoglobulin G detection. The assay used phenolphthalein monophosphate as the substrate, which, when developed, can easily be read visually. Rubestat compared very favorably to hemagglutination inhibition and commercial enzyme-linked immunosorbent assays in its ability to determine immune status. Rubestat demonstrated greater than 97% specificity, sensitivity, and accuracy as compared with other methodologies at 10 different laboratories. The Rubestat index values were precise, with coefficients of variation for intra- and interassay variation of less than 10%. Mean index values had a linear correlation with hemagglutination inhibition titers (r2 greater than 0.97). A population distribution of index values illustrated two distinct bell-shaped curves representing the positive and negative populations. Studies of acute and convalescent serum pairs showed Rubestat to be as accurate as hemagglutination inhibition in determining seroconversion.