Biomedical subjects
Publications and source records attributed to R B Pollard.
Viral resistance limits the value of drugs that act through competitive inhibition of HIV reverse transcriptase (RT). Thus, the emphasis of current research is on agents with other mechanisms. The possibilities include noncompetitive RT inhibitors, drugs against an entirely different enzyme--HIV proteinase, and measures to enhance immunity, either globally or against HIV specifically.
The antiviral effect of Keishi-ni-eppi-ichi-to (TJS-064), a traditional Chinese herbal medicine, was investigated in mice infected with influenza A2(H2N2) virus. When mice exposed to a 5 LD50 dose of the virus were treated orally with a 70 mg/kg dose of TJS-064 1 day before and 1 day and 4 days after the infection, 100% survived over a 25-day experimental period. At the end of this period all the control mice, treated with saline alone, had died; their mean survival time in days (MSD) was 11.2 days. When mice infected with a 10 LD50 dose of the virus were treated with TJS-064, the MSD was > 17.4 days and there was a 50% survival rate, while the control group had a MSD of 8.7 days and a 0% survival rate. No significant antiviral effect of TJS-064 was observed when the agent was administered orally to mice infected with a 100 LD50 or larger dose of influenza virus. Pulmonary consolidations, virus titers in lung tissues and HAI titers in sera of infected mice treated with TJS-064 were all significantly lower than those of infected mice treated with saline. Interferon activities were detected in sera of mice treated with the agent at a dose of 100 mg/kg orally. Since viricidal and viristatic activities of the agent against influenza virus were not demonstrated, the antiviral effects of TJS-064 may be expressed through the host's antiviral functions including interferon production.
The antiviral activity of Shigyaku-to (TJS-109), a traditional Chinese herbal medicine, was investigated in mice infected with herpes simplex virus type 1 (HSV-1). TJS-109 is a combination of the medicinal plant extracts from Zingiberis siccatum rhizoma, Aconiti tuber and Glycyrrhizae radix in a specific proportion. Mice infected with a 10 LD50 dose of HSV-1 were treated with TJS-109 orally at doses of 1.25 to 20 mg/kg 2 days before, and 1 and 4 days after the infection. The treated groups had 80% (1.25 mg/kg), 40% (5 mg/kg) and 23% (20 mg/kg) mortality rates 25 days after the infection as compared with a 100% mortality rate in control mice treated with saline. When HSV-1 infected mice (recipients) received CD8+ T cell fractions derived from spleens of mice treated with TJS-109 (donors), 70% of recipients survived, as compared with 0% survivors in the groups of mice treated with saline, B cell fractions, CD4+ T cell fractions or macrophage-enriched fractions prepared from the same donors. TJS-109 did not show any virucidal activities against HSV-1 or any virostatic activities on the growth of HSV-1 in Vero cells. These results suggest that TJS-109 protected mice exposed to lethal amounts of HSV-1 through the activation of CD8+ T cells.
The effect of Z-100, a lipid-arabinomannan extracted from Mycobacterium tuberculosis strain Aoyama B, was investigated on the resistance of thermally injured mice (TI-mice) to herpes simplex virus type 1 (HSV) infections. The susceptibility of TI mice to infection was about 100 times greater than it was in normal mice (N mice). However, the increased susceptibility of TI mice to infection was effectively counteracted to the levels observed in N mice when treated with Z-100 (10 mg/kg i.p.; 1, 3 and 5 days after thermal injury). Adoptive transfer of burn-associated CD8+ CD11b+ TCR gamma/delta + suppressor T (BAST) cells, prepared from TI mice, increased the susceptibility of N mice to infection by HSV, while the susceptibility of N mice, inoculated with the CD8+ T-cell fraction prepared from Z-100-treated TI mice (ZTC), to infection was not changed. In addition, the suppressor cell activity of BAST cells was not demonstrated when they were assayed in vitro in the presence of anti-IL-4 monoclonal antibody (mAb). BAST cells released IL-4 into their culture fluids without stimulation. The suppressor cell activity of ZTC and IL-4 production by ZTC were minimal. These results suggest that Z-100 may improve the resistance of TI mice to HSV infection through the regulation of BAST cells and/or the release of IL-4 from these cells.
Effect of Kanzo-bushi-to (TJS-038), one of the Japanese Kampo medicines originating from traditional Chinese herbal medicine, on the activity of a clone of burn-associated CD8+ suppressor T cells (T6S cells) was investigated. The suppressor cell activity of T6S cells in the mixed lymphocyte-tumor cell reaction (MLTR) was not changed when various concentrations of TJS-038 were added to the MLTR. The suppressor cell activity of T6S cells was greatly inhibited by CD3+ CD4+ CD28+ TCR alpha/beta+ Vicia villosa lectin-adherent cells (TJS cells) purified from spleen cells of mice treated with TJS-038 at an oral dose of 50 mg/kg. The suppressor cell activity of T6S cells was not demonstrated in the MLTR assay when anti-IL-4 monoclonal antibody was added. Also the production of IL-4 from T6S cells was not detected, when they were co-cultured with TJS cells. These results suggest that TJS-038 stimulates the generation of contrasuppressor T cells (TJS cells) that inhibit the suppressor cell activity of T6S cells and the production of IL-4 from T6S cells.
CMV has been reported to be associated with a DNA polymerase activity (DPA). In this communication its purification, characterization and potential diagnostic value were examined. CMV DNA polymerase was prepared from cell free supernatants of CMV (AD 169) infected cultures. Separation and purification of the enzyme was accomplished by column chromatography of the purified, lysed virus. CMV DPA was measured on an oligo (dT)-poly (dA) template primer. SDS-PAGE and western blot analysis under reducing conditions using an anti-CMV early antibody showed an 80 kDa protein band that was associated with the peak of polymerase activity. However, CMV isolates and CMV from urines from CMV retinitis patients immunoblotted by the same Ab revealed 140 kDa and 80 kDa bands under non-reducing and reducing conditions respectively, the latter was also associated with a 58 kDa band. The diagnostic value of the CMV associated DAP was tested using CMV positive urines. The latter demonstrated high PAA-sensitive DPA activity, compared to normal, HSV positive urines and urines from HBSAg positive patients. Taken collectively, these findings indicate the potential usefulness of CMV-associated DNA polymerase activity in the diagnosis and follow-up of patients with CMV-related illnesses.
Human CMV causes a number of diseases that cause considerable morbidity and that can be life-threatening in immunocompromised patients, particularly those with AIDS. Ganciclovir (GCV) and Foscarnet (PFA) are currently the drugs of choice for management of CMV disease. Both are not without side effects and have a relatively narrow margin of safety. In this report the effects of a human IgG1 neutralizing monoclonal antibody MSL-109 (MSL, Sandoz Pharmaceuticals) on CMV replication was examined both alone or in combination with either GCV or PFA. Human embryonic lung fibroblasts were infected with CMV strain AD169 with a multiplicity of infection of 3 plaque forming units/cell for 1 h. Prior to infection the virus was incubated for 30 min at 37 degrees C with serial concentrations of the MSL Ab (0.1-3.0 micrograms/ml). Concentrations of GCV (0.3 to 30 microM) or PFA (50-400 microM) were added to CMV-infected cells that had been either previously incubated with MSL or not. Four days after infection CMV replication was measured by DNA/DNA probe hybridization using the Hybriwix system. MSL in combination with GCV had an additive effect that was observed at concentrations of GCV of 3-10 microM and MSL of 1-10 micrograms/ml. On the other hand, MSL (3-10 micrograms/ml) together with PFA (100-400 microM) produced a synergistic effect on CMV replication. The data suggest that MSL at doses achievable in humans, enhanced GCV- and PFA-induced antiviral effect in a dose-dependent manner and that the combination might be clinically useful in the treatment of CMV disease.
The protective effect of Kanzo-bushi-to (TJS-038) was investigated on the opportunistic infection of herpes simplex virus type 1 (HSV) in thermally injured mice (TI-Mice). We have previously reported that TI-Mice were approximately 100 times more susceptible to HSV infection than normal mice (N-Mice) and that CD8+ suppressor T (ST)-cells induced by burn injury were involved in causing this increased susceptibility of TI-Mice. Increased susceptibility of TI-Mice to the infection was reversed to the levels observed in N-Mice when TI-Mice were treated intraperitoneally with TJS-038 at a dose of 5 mg/kg 1 and 4 days after thermal injury. The activity of ST-cells was greatly decreased in TI-Mice treated with TJS-038. The generation of Vicia villosa lectin-adherent CD4+ CD28+ TCR-alpha/beta+ contrasuppressor T (Contra-ST)-cells associated with the appearance of ST-cells was expanded and occurred earlier in spleens of TJS-038-treated TI-Mice as compared with that of untreated TI-Mice. The improved resistance of TJS-038-treated TI-Mice to the infection was transferred to untreated TI-Mice by adoptive transfer of Contra-ST-cells prepared from TJS-038-treated TI-Mice. These results suggest that TJS-038 may restore the resistance of TI-Mice to the HSV infection through the expanded generation of Contra-ST-cells.
Protein Kinase C (PKC) and Ca++ are both involved in the chain of events leading to T-cell activation. An impairment of the immune response is characteristic of T cells obtained from patients with HIV infection. In this report, the involvement of PKC and Ca++ in HIV-mediated cellular hyporesponsiveness was examined. Infection of peripheral blood mononuclear cells (PBMC)s from HIV-seronegative normal donors with HIV strain HTLV IIIB, or two fresh patient isolates produced a 1.4-, 10.7-, and 11.4-fold enhancement in PKC activity at 1 hr postinfection (PI) and a 1.8-, 2.3-, and 3.8-fold enhancement at 12 hr PI, respectively. A marked decrease of PKC content, as determined by Western Blot analysis, was observed in HIV-infected cells by Day 4 and 7 PI compared with mock-infected control cells. Furthermore, PKC synthesis was also inhibited in cells from immunosuppressed AIDS patients. PKC activity of PBMCs from HIV-infected patients did not change in response to 1 microM of phorbal myristate acetate (PMA). In contrast, the same dose enhanced the activity by 50%-100% in PBMCs from normal HIV-seronegative donors. A 40%, 50%, and 125% increase in intracellular free Ca++ in response to HIV infection was observed 12 hr PI in MT4, JURKAT, and PBMCs, respectively. However, the increase in intracellular free Ca++ in HIV-infected PBMCs obtained from normal donors in response to PHA was 56% and 17% compared with an increase of 100% and 120% in mock infected cells at 12 hr and 1 week PI, respectively. Comparing the Ca++ response to PHA in PBMCs from HIV-infected patients showed that patients with < 250 absolute T4 cells/mm3 had an impaired Ca++ response. These data suggest that there is a relationship between intracellular free Ca++ and PKC and HIV-induced T-cell hyporesponsiveness.
OBJECTIVE: To investigate the likelihood of patients who have human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS) being parents and to identify concerns of these parents about their children. DESIGN: A survey was conducted of parental status, demographics, perceptions of social/emotional needs of self and one's children. Responses were analyzed for demographic differences. PARTICIPANTS: A total of 242 patients from the university HIV/AIDS clinics completed the survey. MAIN OUTCOME MEASURES: Parental status, number, and ages of children, parental concerns about their children related to their own HIV/AIDS: RESULTS: Nearly one third (31.8%) of the sample of HIV/AIDS patients were parents, and three fourths (76%) of the female patients were mothers. Slightly more than one third of these were married, and these were not predominantly families who also had infected children. The percentage of women in the parent subsample (40.8%) was higher than the percentage of women in the overall patient sample (16.7%). Only half of the parents reported that their children > 4 years of age knew of their diagnosis. Two thirds of the parents reported they believed their children did not need to talk to someone about their parent's health, and nearly half of the parents reported that they did not need help dealing with their children concerning issues related to AIDS. CONCLUSION: The percentage of HIV/AIDS patients who are parents is high, and parental status and emotional needs of parents and their children will likely become an increasingly important issue. Many questions are raised by our findings. Should we be concerned that many parents have been unable to talk to their children about their own health? Should we help parents acknowledge that their children may need some outside help to cope?
A population of B cells that regulates the immune response was demonstrated in splenic mononuclear cells (SMNC) from mice infected with herpes simplex virus type 1 (HSV). SMNC, obtained from mice 3 days after HSV infection at a dose of 100 LD50, exhibited a reduction in the proliferative response of naive SMNC stimulated with various lectins or allogeneic lymphocytes in a 5-day mixed lymphocyte reaction. Cocultivation of naive SMNC with phagocytic cell-free SMNC (M phi-MNC) from infected mice resulted in the inhibition of lymphocytic blast transformation stimulated with various lectins. These cells were characterized as nylon-wool adherent cells that were eliminated by treatment with anti-Ig antiserum, but not anti-Thy 1.2 monoclonal antibody or anti-asialo GM1 antiserum, followed by complement treatment. In addition, the suppressor cell activity was not demonstrated in M phi-MNC obtained from HSV-infected CBA/CaHN-xid/J mice, which contain a congenital B-cell deficiency. These results suggest that, in addition to suppressor T cells, a population of B cells, which can inhibit lymphocyte proliferations upon stimulation with lectins and alloantigens, might be generated in spleens of mice following HSV infections.
Immunomodulatory effects of oily formulated SMANCS, a polymer conjugated derivative of the proteinaceous antibiotic neocarzinostatin, after oral administrations to mice was investigated. The oral administrations of SMANCS, dissolved in medium-chain triglyceride containing phosphatidylcholine and polyglycerine dioleate (oily SMANCS), resulted in: (1) augmentation of NK cell activity in naive mice, (2) activation of macrophage cytostasis in naive mice, (3) enhancement of the generation of cytotoxic T-lymphocytes in mice immunized with allogeneic lymphocytes, (4) production of circulating interferon in naive mice, and (5) increase of delayed-type hypersensitivity response in mice immunized with sheep red blood cells. The degree of immunomodulation orally stimulated with oily SMANCS was similar to that of the immunomodulation induced by i.v. or i.p. administrations of aqueous formulated SMANCS (aqueous SMANCS). Although SMANCS is a protein drug that may be digestable by various enzymes present in the stomach, in the present study immunomodulating activities of SMANCS were clearly demonstrated when an oily formulation of the compound was administered to mice orally. Since aqueous SMANCS administered parenterally and oily SMANCS administered orally exhibit the same immunomodulatory activities and the former has demonstrated antitumor activity in man and animals, the latter may possess this same antitumor activity.
The inhibitory effect of glycyrrhizin (GR), an anti-inflammatory Chinese herbal drug extracted from licorice roots, on the generation of suppressor T cells in thermally injured mice (TI-mice) was investigated. The burn-associated suppressor T cell (BTs cell) activity was demonstrated in splenic mononuclear cells (SMNC) from mice 2 to 8 days after thermal injury when suppressor cell activity was assayed in a one-way mixed lymphocyte reaction. However, when TI-mice were treated with GR, SMNC harvested 6 days after thermal injury showed no suppressor cell activity. This activity of GR demonstrated a dose-response effect, with a dose of 10 mg/kg exhibiting peak levels of the activity. Since GR had no direct inactivating activities against BTs cells in vitro, the inhibitory effect of SMNC, derived from TI-mice treated with GR, on the activity of BTs cells was examined in the same mixed lymphocyte reaction system, and the results showed that the SMNC from GR-treated mice 6 days after thermal injury counteracted the activity of BTs cells. The type of cell responsible for this inhibition of BTs cell activities was a CD3+, L3T4+, Vicia villosa lectin-adherent T cell with the same phenotypic properties previously exhibited by contrasuppressor cells. These results suggest that GR may regulate the generation of BTs cells through the induction of contrasuppressor cells. Since there are many reports describing septic infections due to the appearance of BTs cells in postburn patients, it may be possible to apply GR or blood preparations containing contrasuppressor cell populations induced by GR in healthy volunteers into immunosuppressed burn patients to avoid infections.
This prospective, randomized, multicenter, controlled trial was designed to evaluate the efficacy and safety of intravenous ganciclovir for the treatment of peripheral cytomegalovirus (CMV) retinitis in patients with AIDS. Patients were randomly assigned to receive either immediate treatment, intravenous ganciclovir, 5 mg/kg twice daily for 14 days followed by 5 mg/kg once daily for 14 weeks, or deferred treatment. Patients randomized to deferred treatment whose retinitis progressed were offered ganciclovir. Of the 22 patients randomized to deferred treatment who were included in the final analysis, 20 were found to have progressive CMV retinitis compared with 10 of the 13 randomized to immediate treatment. The median time to progression in the deferred treatment group, as determined by a masked fundus photography reading center, was 13.5 days compared with 49.5 days in the immediate treatment group (mean +/- SD, 19.3 +/- 4.1 vs. 66.4 +/- 14.0; P = .001, log rank test). These data indicate that ganciclovir delays the progression of CMV peripheral retinitis in persons with AIDS.
We reviewed the hospital charts of 17 patients with AIDS and Clostridium difficile diarrhea to determine antibiotic use before C. difficile infection, methods of treatment for C. difficile diarrhea, and response of diarrhea to treatment. Left shift and total white blood cell count before and after treatment for C. difficile were also determined. Non-HIV-infected patients with C. difficile diarrhea served as controls. In the patients with AIDS, resolution of diarrhea was noted in 15 (88%) patients. In 25 (76%) control patients, diarrhea resolved with treatment. The patients with AIDS also had a significant decrease (p < 0.05) in left shift in white blood cell count with treatment; the controls did not. Our study therefore suggests that C. difficile diarrhea is at least as likely to resolve with antibiotic therapy in patients with AIDS as it is in those with the non-AIDS-related disorder. We also found that patients with AIDS and C. difficile diarrhea are more likely than patients without AIDS to have a decreased left shift in white blood cell count after antibiotic therapy.
Cytomegalovirus (CMV) infection constitutes a serious threat to patients with acquired immune deficiency syndrome. Recently we reported that human immunodeficiency virus (HIV) infection of CD4+ cells was associated with sustained elevation of cellular levels of cAMP. Moreover, cyclic nucleotide modulators enhanced HIV replication by increasing intracellular levels of cAMP. In this study, the effect of CMV on HIV replication in CMV/HIV mixed infection and its relationship to cAMP were examined. MT-4 cells, CMV strain AD169, and HIV strain IIIB were used. Optimal enhancement (4.4-fold increase) of HIV replication was observed when MT-4 cells were infected with CMV at Day 0 followed by HIV on Day 4 after infection, as determined by reverse transcriptase activity on Day 11 after infection. cAMP (measured by radioimmunoassay) levels in cells infected with CMV alone, HIV alone, or CMV/HIV together were 2-, 3-, and 5-fold above untreated cells, respectively. CMV also enhanced the replication of UV-irradiated HIV 4-fold and this was associated with a 2-fold increase in cAMP as well. Moreover, UV-irradiated CMV enhanced HIV replication 8.8-fold. The same dose of viable and UV-irradiated CMV used in the above experiments increased protein kinase C activity in these cells 3.0- and 8.0-fold, respectively. These findings might suggest that cAMP and protein kinase C are involved in CMV enhancement of HIV replication. These findings may have relevance to the identification of novel target sites for development of antiviral therapeutics.
The protective effects of combination therapy utilizing recombinant human TNF-alpha (rTNF-alpha) and natural murine interferon-beta (IFN-beta) in mice infected with herpes simplex virus type 1 (HSV-1) was investigated. Mice treated with rTNF-alpha alone at all of the doses tested (a single i.v. administration, 2.3-2,300 micrograms/kg; multiple i.p. administrations 0.4-250 micrograms/kg) as well as mice that received IFN-beta alone at doses of 16 x 10(4) U/kg or less resulted in a 0% survival rate. Combination therapy consisting of a single administration of rTNF-alpha (230 and 23 micrograms/kg) and multiple administrations of IFN-beta (4 x 10(4) U/kg) resulted in a 40% and 60% survival rate. Multiple treatments of infected mice with rTNF-alpha (50 and 10 micrograms/kg) in combination with IFN-beta (4 x 10(4) U/kg) resulted in 50% and 70% survival rates, respectively. These results suggest that the combination therapy of rTNF and natural murine IFN-beta produce synergistic protective effects in mice infected with a lethal amount of HSV-1.
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