ABSTRACT
OBJECTIVE: To assess the safety of live, attenuated influenza vaccine (LAIV) administered to relatively asymptomatic or mildly symptomatic HIV-infected children and non-HIV-infected children. METHODS: Twenty-five non-HIV and 24 HIV-infected children (CDC Class N or A1,2) were enrolled into this double blind, placebo-controlled study. Children were randomized within each HIV status group to one of two dosing regimens: Regimen 1, Dose 1 = LAIV, Dose 2 = placebo, Dose 3 = LAIV; or Regimen 2, Dose 1 = placebo, Dose 2 = LAIV, Dose 3 = LAIV. Study doses were separated by 28 to 35 days. Reactogenicity events within 10 days and adverse events within 28 to 35 days after each study dose were recorded. Blood HIV RNA concentrations, CD4 counts and CD4% were measured throughout the study on HIV-infected children. Quantitative influenza cultures were performed on nasal aspirates collected periodically from all children up to 28 to 35 days after each study dose. Influenza isolates were assessed for retention of the temperature-sensitive phenotype. Serum influenza HAI antibodies were measured before and after each LAIV vaccination. RESULTS: No significant differences were found in rates of reactogenicity events and vaccine-related adverse events after placebo or the first dose of LAIV within each HIV status group, nor were differences found between HIV-infected and HIV-uninfected children after each dose of LAIV. Overall none of the HIV-infected children experienced a significant LAIV-related serious adverse event or influenza-like illness, making the one sided 95% CI of such a serious event occurring after LAIV 0 to 12%. No significant changes in geometric mean HIV RNA concentrations, CD4 counts or CD4% or prolonged or increased quantity of LAIV virus shedding occurred in HIV-infected children after receiving either dose of LAIV. All recovered influenza isolates retained the temperature-sensitive phenotype. After two doses of LAIV, 83% of the non-HIV-infected and 77% of the HIV-infected children had a > or = 4-fold rise in influenza antibody to at least one of the three LAIV strains. CONCLUSION: If relatively healthy HIV-infected children become exposed to LAIV inadvertently, then serious adverse outcomes would not be expected to occur frequently.
Subject(s)
HIV Infections/immunology , Influenza Vaccines/adverse effects , Influenza Vaccines/immunology , Vaccination , Vaccines, Attenuated/adverse effects , Vaccines, Attenuated/immunology , Child , Child, Preschool , Cold Temperature , Double-Blind Method , HIV Infections/virology , HIV-1/physiology , Hemagglutination Inhibition Tests , Humans , Infant , Influenza A virus/immunology , Influenza A virus/isolation & purification , Influenza B virus/immunology , Influenza B virus/isolation & purification , Influenza Vaccines/administration & dosage , Influenza, Human/prevention & control , Vaccines, Attenuated/administration & dosage , Virus SheddingABSTRACT
The authors provide an analysis of data from a two-year (1996-1998), multicenter (ten US cities), double-blinded, placebo-controlled influenza vaccine trial in children. The vaccine was the trivalent cold-adapted influenza vaccine. Estimates are made of the vaccine efficacy for susceptibility to culture-confirmed influenza (VE(S)) while taking inter-center variability in the risk of infection into account. Our overall estimate of VE(S) against influenza is 0.92 (95% confidence interval (CI) 0.89-0.94). In addition, for the second year, although the vaccine contained antigen for A/Wuhan-like (H3N2), the estimated VE(S) for epidemic variant A/Sydney-like (H3N2) was 0.89 (95% CI 0.81-0.94). Thus, the vaccine showed a high degree of protection against a variant not closely matched to the vaccine antigen. With regard to natural immunity, an influenza A infection in the first year reduces the estimated risk of an influenza A infection in the second year by a factor of 0.88 (95% CI 0.21-0.98). When comparing year 1 to year 2, there is a negative correlation of -0.50 in the center-specific attack rates in the placebo groups. This is consistent with the theory that natural immunity provides overall community protection to children. The authors argue that mass vaccination of 70% of the children with the cold-adapted influenza vaccine could provide substantial protection to the community at large.
Subject(s)
Influenza Vaccines/therapeutic use , Influenza, Human/prevention & control , Child, Preschool , Double-Blind Method , Humans , Infant , Influenza A virus/immunology , Influenza B virus/immunology , Influenza Vaccines/immunology , Influenza, Human/immunology , Placebo Effect , Treatment Outcome , Vaccines, Attenuated/immunology , Vaccines, Attenuated/therapeutic useABSTRACT
An investigational live influenza virus vaccine, FluMist, contains three cold-adapted H1N1, H3N2, and B influenza viruses. The vaccine viruses are 6/2 reassortants, in which the hemagglutinin (HA) and neuraminidase (NA) genes are derived from the circulating wild-type viruses and the remaining six genes are derived from the cold-adapted master donor strains. The six genes from the cold-adapted master donor strains ensure the attenuation, and the HA and NA genes from the wild-type viruses confer the ability to induce protective immunity against contemporary influenza strains. The genotypic stability of this vaccine was studied by employing clinical samples collected during an efficacy trial. Viruses present in the nasal and throat swab specimens and in supernatants after culturing the specimens were detected and subtyped by multiplex reverse transcriptase (RT)-PCR. Complete genotypes of these detected viruses were determined by a combination of RT-PCR and restriction fragment length polymorphism, multiplex RT-PCR and fluorescent single-strand conformation polymorphism, and nucleic acid sequencing analysis. The FluMist vaccine appeared to be genotypically stable after replication in the human host. All viruses detected during the 2-week postvaccination period were shed vaccine viruses and had maintained the 6/2 genotype.
Subject(s)
Influenza A virus/genetics , Influenza B virus/genetics , Influenza Vaccines , Vaccination , Cold Temperature , Double-Blind Method , Genotype , Humans , Influenza A virus/classification , Influenza A virus/immunology , Influenza A virus/isolation & purification , Influenza B virus/classification , Influenza B virus/immunology , Influenza B virus/isolation & purification , Influenza Vaccines/genetics , Influenza, Human/prevention & control , Nose/virology , Pharynx/virology , Polymorphism, Restriction Fragment Length , Polymorphism, Single-Stranded Conformational , Reverse Transcriptase Polymerase Chain Reaction , Sequence Analysis, DNAABSTRACT
Fifty-seven human immunodeficiency virus (HIV)-infected (CDC class A1-2) and 54 non-HIV-infected adults, not prescreened for influenza susceptibility, were randomized to receive trivalent live attenuated influenza vaccine (LAIV) or placebo intranasally. LAIV was safe and well tolerated with no serious adverse events attributable to vaccine. Reactogenicity rates were similar in LAIV and placebo recipients except that runny nose/nasal congestion was significantly more common in LAIV recipients regardless of HIV status. No prolonged shedding of LAIV was observed in HIV-infected participants. HIV RNA levels were not increased and CD4 counts were not decreased in HIV-infected LAIV recipients compared with placebo recipients after immunization. Shedding of LAIV and increases in antibody titers were infrequent, consistent with prior experience in unscreened adults. The data suggest that inadvertent vaccination with LAIV in relatively asymptomatic HIV-infected adults would not be associated with frequent significant adverse events.
Subject(s)
HIV Infections/immunology , Influenza A virus/immunology , Influenza B virus/immunology , Influenza Vaccines/adverse effects , Influenza Vaccines/immunology , Adult , Antibodies, Viral/blood , Cold Temperature , Female , HIV Infections/virology , HIV-1/physiology , Hemagglutination Inhibition Tests , Humans , Influenza A virus/isolation & purification , Influenza B virus/isolation & purification , Influenza Vaccines/administration & dosage , Male , RNA, Viral/blood , Vaccination , Vaccines, Attenuated/administration & dosage , Vaccines, Attenuated/adverse effects , Vaccines, Attenuated/immunology , Virus SheddingABSTRACT
Twenty-six human immunodeficiency virus (HIV)-infected pregnant women participated in a placebo-controlled study of immunogenicity and safety of multiple doses of MN rgp120 vaccine over the last half of pregnancy. The women had CD4 lymphocyte counts>400/mm3, no AIDS-defining illness and normal pregnancies. Vaccination was well tolerated, with no significant local or systemic reactions in the women and no adverse outcomes in the infants attributable to the vaccine. Vaccination did not alter plasma RNA reverse transcriptase-polymerase chain reaction copy number; moreover, immunization was not associated with changes in CD4 counts or HIV binding and neutralization antibody titers. Infants were followed up until 18 months of age. Five of 26 infants (19%) were HIV infected, with infection occurring in children of both vaccinated and placebo women. Analysis of factors that influence transmission did not disclose associations with immunization status, viral load, CD4 count, or maternal viral neutralization titers.
Subject(s)
AIDS Vaccines/adverse effects , CD4 Lymphocyte Count , HIV Envelope Protein gp120/adverse effects , HIV Infections/therapy , Infectious Disease Transmission, Vertical/prevention & control , Pregnancy Complications, Infectious/immunology , Vaccines, Synthetic/adverse effects , Adolescent , Adult , Antibody-Dependent Cell Cytotoxicity , Female , Follow-Up Studies , HIV Antibodies/blood , HIV Infections/immunology , HIV Infections/prevention & control , Humans , Infant, Newborn , Placebos , Pregnancy , Pregnancy Complications, Infectious/virology , Safety , Time FactorsABSTRACT
Among 2099 uninfected subjects in phase I and II trials of candidate AIDS vaccines, 23 were diagnosed with intercurrent human immunodeficiency virus type 1 (HIV-1) infection. High-risk sexual exposures accounted for 17 infections, and intravenous drug use accounted for 6. Four subjects received placebo, 13 received a complete immunization schedule (> or = 3 injections), and 6 were partially immunized (< or = 2 injections). There was no significant difference between vaccine recipients and control groups in incidence of HIV-1 infection, virus load, CD4 lymphocyte count, or V3 loop amino acid sequence. In summary, 19 vaccinated subjects acquired HIV-1 infection during phase I and II trials, indicating that immunization with the products described is < 100% effective in preventing or rapidly clearing infection. Laboratory analysis suggested that vaccine-induced immune responses did not significantly affect the genotypic or phenotypic characteristics of transmitted virus or the early clinical course of HIV-1 infection.
Subject(s)
AIDS Vaccines/therapeutic use , HIV Infections/diagnosis , HIV Infections/prevention & control , HIV-1 , Adult , Amino Acid Sequence , CD4 Lymphocyte Count , Female , HIV Antibodies/analysis , HIV Envelope Protein gp120/analysis , HIV Envelope Protein gp120/genetics , HIV Infections/therapy , Humans , Immunity, Active , Incidence , Male , Middle Aged , Molecular Sequence Data , Neutralization Tests , Peptide Fragments/analysis , Peptide Fragments/genetics , Risk-Taking , Sequence Analysis , Substance Abuse, Intravenous , Viral LoadABSTRACT
OBJECTIVE: To evaluate the safety and immunogenicity of recombinant glycoprotein (rgp) 120, a candidate vaccine for the human immunodeficiency virus (HIV), formulated with a novel adjuvant, MF59, with or without a biological response modifier, MTP-PE. DESIGN: Multicenter, double-blind, randomized trial. SETTING: University medical centers. PARTICIPANTS: 49 healthy, HIV-seronegative volunteers 18 to 60 years of age who were at low risk for HIV type 1 (HIV-1) infection. INTERVENTIONS: In part A of the study, 32 participants were randomly assigned to receive either 15 micrograms of rgp 120 in MF59, 15 micrograms of rgp 120 in MF59 plus 50 micrograms of MTP-PE, 50 micrograms of rgp 120 in MF59, or 50 micrograms of rgp 120 in MF59 plus 50 micrograms of MTP-PE. Participants were vaccinated at 0, 1, 6, and 12 to 18 months. In part B, 17 participants were randomly assigned to receive five monthly injections of either 50 micrograms of rgp 120 in MF59 or MF59 alone followed by a booster injection at 12 to 18 months. MAIN OUTCOME MEASURES: Local and systemic reactions; laboratory measures of hepatic, renal, immunologic, and bone marrow toxicity; and HIV-specific serologic and cell-mediated immune responses. RESULTS: 13 patients in part A received 50-micrograms doses of rgp 120; type-specific neutralizing antibody responses against the SF-2 strain of HIV-1 (HIV-1/SF-2) were induced in all 13. Nine of the 13 had crossreactive neutralizing activity against the MN strain of HIV-1 (HIV-1/MN), and 2 had crossreactive neutralizing activity against the IIIB strain of HIV-1 (HIV-1/IIIB). Twelve patients had typespecific fusion inhibition activity; only 1 had crossreactive fusion inhibition activity against HIV-1/MN. The monthly vaccination schedule used in part B resulted in decreased antibody titers, indicating that a rest period in the schedule is necessary for maximal immunogenicity. Lymphoproliferative responses against gp120 were induced in all vaccine recipients. The stimulation index to gp120 was persistently greater than 15 for 6 months after the last booster vaccination was given. CD8+ cytotoxic T-lymphocyte activity was detected in 1 of the 11 participants tested. Vaccine that contained MTP-PE caused a greater number of moderate or severe local and systemic reactions (of 16 participants, 4 had local reactions and 13 had systemic reactions) than did vaccine formulated with MF59 alone (of 16 participants, 7 had local reactions [P < 0.01] and 0 had systemic reactions [P < 0.001]). CONCLUSIONS: The SF-2 rgp120 vaccine is safe and immunogenic. Three vaccinations with rgp120 in MF59 can induce type-specific and crossreactive neutralizing antibody against B-subtype laboratory strains of HIV-1. Human immunodeficiency virus-specific lymphoproliferative responses were induced in all vaccinated participants, and CD8+ cytotoxic T-lymphocyte activity was shown in one participant. A trend toward the augmentation of lymphoproliferative and humoral responses by MTP-PE was seen in the participants receiving 15 micrograms of rgp120. However, MTP-PE caused a statistically significant increase in the incidence of local and systemic side effects, which was felt to outweigh the small increase in immunogenicity provided by this biological response modifier in an otherwise well-tolerated vaccine.
Subject(s)
AIDS Vaccines/adverse effects , AIDS Vaccines/immunology , HIV Antibodies/blood , HIV-1/immunology , Adult , Double-Blind Method , Female , HIV Envelope Protein gp120/adverse effects , HIV Envelope Protein gp120/immunology , HIV Seronegativity , Humans , Immunization Schedule , Male , Middle Aged , Recombinant Proteins/adverse effects , Recombinant Proteins/immunology , Reference Values , T-Lymphocytes/immunologyABSTRACT
We investigated the safety and immunogenicity of a candidate HIV-1 vaccine, Env 2-3 (Chiron Biocine Co.), in combination with an adjuvant emulsion, MF59, with or without an additional immune modulator, MTP-PE 78 healthy HIV-1-seronegative adults. Sixteen subjects participated in a dose escalation study of MTP-PE in MF59 without Env 2-3, given at 0 and 1 months; 48 subjects participated in a study of a fixed dose of 30 micrograms of Env 2-3 in MF59 with increasing doses of MTP-PE (0, 5, 10, 25, 50, and 100 micrograms), and 14 subjects participated in a study of 100 micrograms of Env 2-3 in MF59 without MTP-PE. Subjects were assigned to study groups under a randomized, double-blind allocation. Subjects received immunization at 0, 1, and 6 months, and had the option of receiving a fourth dose at 12-18 months. Env 2-3 in MTP-PE/MF59 was associated with significant reactogenicity, in that severe, although self-limited systemic and/or local reactions occurred in 15 of 30 vaccinees. In contrast, Env 2-3 in MF59 without MTP-PE was relatively well tolerated, and severe local and/or systemic reactions occurred in only 2 of 18 subjects. Env 2-3 stimulated serum antibodies to HIV-1 envelope protein (gp120) as detected by Western blot in 39 of 43 subjects and to HIV-1 virus lysate by EIA in 28 of 43 subjects after three injections. The majority of subjects also developed EIA antibodies to recombinant gp120 (SF-2), gp120 (LAI), and V3 peptide (SF-2). Neutralizing antibodies to the homologous SF-2 strain developed in 30 of 43 and 27 of 34 subjects, and fusion inhibition antibodies in 25 of 43 and 15 of 36 subjects after three and four injections, respectively. Lymphoproliferative responses to the immunogen, Env 2-3 were observed in over 80% of the vaccinees examined, and CD4+ cytotoxic T cell activity directed against HIV-1 was noted transiently in 2 of 20 vaccinees. Addition of MTP-PE to Env 2-3 or increasing the dose of Env 2-3 from 30 to 100 micrograms did not augment immunogenicity. Env 2-3 in MF59 was well tolerated and immunogenic in HIV-1-seronegative individuals. The addition of MTP-PE significantly increased reactogenicity, but had little, if any, effect on immunogenicity.
Subject(s)
AIDS Vaccines/administration & dosage , Adjuvants, Immunologic/administration & dosage , HIV Envelope Protein gp120/immunology , HIV Infections/prevention & control , HIV-1/immunology , Polysorbates/administration & dosage , Squalene/administration & dosage , AIDS Vaccines/immunology , Adolescent , Adult , Amino Acid Sequence , Cells, Cultured , Consumer Product Safety , Dose-Response Relationship, Immunologic , Double-Blind Method , Female , HIV Antibodies/blood , HIV Infections/immunology , Humans , Leukocytes, Mononuclear/cytology , Leukocytes, Mononuclear/immunology , Male , Middle Aged , Molecular Sequence Data , Squalene/immunology , T-Lymphocytes, Cytotoxic/immunology , Vaccines, Synthetic/administration & dosage , Vaccines, Synthetic/immunologyABSTRACT
A phase I double-blind trial was done to examine the safety and immunogenicity of a prototype synthetic human immunodeficiency virus type 1 MN strain (HIV-1MN) third variable region domain (V3) branched peptide vaccine in HIV-1-uninfected healthy adult volunteers. Subjects were randomly assigned to receive 20, 100, or 500 micrograms of vaccine or alum adjuvant control on days 0, 28, and 168. The vaccine was well-tolerated and appeared safe. Induction of binding antibody to V3 MN branched peptide was vaccine dose-related and was detectable in 9 of 10 subjects in the highest-vaccine-dose group. HIV-1MN-neutralizing antibody was detected after the third 500-micrograms dose in 8 of 10 subjects at the 90% neutralization end point. V3 MN peptide stimulated lymphocyte proliferation in 15 (75%) of 20 subjects after vaccination. In conclusion, this prototype vaccine was safe and it induced humoral and cell-mediated immune responses.
Subject(s)
AIDS Vaccines/administration & dosage , HIV Antibodies/analysis , HIV Envelope Protein gp120/immunology , HIV-1/immunology , Peptide Fragments/immunology , AIDS Vaccines/adverse effects , AIDS Vaccines/immunology , Adult , Amino Acid Sequence , Dose-Response Relationship, Immunologic , Double-Blind Method , Enzyme-Linked Immunosorbent Assay , Female , HIV Envelope Protein gp120/chemistry , Humans , Lymphocyte Activation , Male , Middle Aged , Molecular Sequence Data , Neutralization Tests , Peptide Fragments/chemistrySubject(s)
AIDS Vaccines/pharmacology , HIV Infections/prevention & control , HIV-1/immunology , Clinical Trials, Phase I as Topic/methods , Female , HIV Infections/complications , HIV Infections/transmission , Humans , Immunization, Passive , Infant, Newborn , Infectious Disease Transmission, Vertical , Pregnancy , Pregnancy Complications, InfectiousABSTRACT
Anticipating the availability of one or more candidate HIV vaccines for efficacy testing in the next few years, public health agencies are now planning for the conduct of large-scale efficacy trials. We expect these trials to be randomized, double-blind, placebo-controlled studies with prevention of infection as the primary goal. We discuss in detail factors that influence sample size. Factors most influential are the incidence rate of HIV infection in the study population and the minimum efficacy at which a vaccine is still considered acceptable. The smaller either of these factors is, the larger the sample size will be. The desire to complete trials quickly, the gradual accrual of benefit from vaccination, the inaccuracies of assays to detect infection, the need to counsel participants to avoid exposure to HIV, and loss to follow-up all tend to drive up sample size. To illustrate, 83 subjects per study arm suffice to detect 90% efficacy in a population with a 7% annual risk of infection. This assumes a 3-year study with accrual completed in 1 year, no loss to follow-up, and Types I and II error rates of 5 and 10%, respectively. In contrast, 4,254 subjects per arm are required to identify a 60% effective vaccine in a population with a 1% annual risk. The study is also shortened to 2 years, assumes a 5% annual loss to follow-up, and supposes that the full benefit of vaccination is achieved in 6 months. The most realistic assumptions indicate that trials are very likely to require several thousand participants. Limitations of the proposed designs are also discussed.
Subject(s)
Clinical Trials as Topic/standards , HIV Infections/prevention & control , HIV Infections/therapy , Viral Vaccines , Female , HIV Infections/diagnosis , Humans , Immunotherapy , Male , Pregnancy , Pregnancy Complications, Infectious/therapy , Research Design , Viral Vaccines/standardsSubject(s)
HIV Infections/prevention & control , Pregnancy Complications, Infectious , Animals , Clinical Trials as Topic , Female , Fetal Diseases/microbiology , Fetal Diseases/prevention & control , HIV/isolation & purification , HIV Infections/congenital , HIV Infections/epidemiology , HIV Infections/therapy , HIV Infections/transmission , Humans , Immunization, Passive , Immunoglobulins, Intravenous/therapeutic use , Infant , Infant, Newborn , Pregnancy , Pregnancy Complications, Infectious/epidemiology , Pregnancy Complications, Infectious/therapy , Vaccination , Zidovudine/therapeutic useABSTRACT
Adjuvant-like arthritis was produced in Sprague-Dawley rats during 14 days of oral administration of 2-amino-5-bromo-6-(3-fluorophenyl)-4(3H)-pyrimidinone. Inflammatory changes about and in the hindlimb joints were similar to those described by Pearson for adjuvant-induced polyarthritis. Lymphoid hyperplasia, elevation of serum IgG levels, and localization of fluorescein-labeled globulin in disseminated inflammatory lesions implicate an immunologic effect. Rats treated twice weekly for 64 days developed lymphocytic thyroiditis and showed less inflammation in the joints.
Subject(s)
Antineoplastic Agents/toxicity , Arthritis, Experimental/pathology , Arthritis/pathology , Cytosine/analogs & derivatives , Administration, Oral , Animals , Antineoplastic Agents/administration & dosage , Arthritis, Experimental/chemically induced , Arthritis, Experimental/metabolism , Body Weight/drug effects , Cytosine/administration & dosage , Cytosine/toxicity , Female , Hindlimb , Immunoglobulin G/analysis , Interferons/blood , Male , Organ Size/drug effects , Rats , Rats, Inbred StrainsABSTRACT
Immunofluorescence was used to characterize the lymphocyte subpopulations of mice treated with six immunomodulatory drugs: hydrocortisone acetate (HCA), corticosterone acetate (corticosterone), cyclophosphamide, cytosine arabinoside (Ara-C), 15(S)-methyl prostaglandin E1 (15(S)-methyl PGE1), and 2-amino-5-bromo-6-phenyl-4-(3H)-pyrimidinone (ABPP). The number of thymus and spleen cells bearing Thy-1, Ig, Lyt-1 and Lyt-2 antigens and the density of the antigens on each cell (IF profiles) were determined. Microscopic examination of cells stained with rhodamine-labeled anti-Lyt-2 and fluorescein-labeled anti-Lyt-1 was used to measure the proportion of Lyt-1+2-, Lyt-1+2+, and Lyt-1-2+ cells in the spleen and thymus of drug-treated animals. The changes in lymphocyte subpopulations were compared with the varied effects of these drugs on antibody formation and graft vs host (GVH) reaction. Three immunosuppressive drugs, HCA, cyclophosphamide, and Ara-C, depleted the thymus of cells expressing a large quantity of Thy-1. The drug-resistant cells were larger and had more Lyt-1 than cells from control animals. HCA treatment depleted the thymus of Lyt-1+2+ cells; the cortisone resistant cells were primarily Lyt-1+2-. Cyclophosphamide and the antiviral immunostimulant, ABPP, caused similar, but less marked, alterations. The proportion of Lyt-1-2+ cells in the thymus was reduced by treatment with all the drugs, but the density of Lyt-2 on the drug-resistant cells was not altered. Treatment with Ara-C or 15(S)-methyl PGE1 produced a very modest evaluation in Lyt-1+2- cells. 15(S)-Methyl PGE1, which suppresses some immuno-inflammatory reactions, had no discernible effect on thymocyte size or the IF profile of Thy-1, Lyt-1, or Lyt-2. In the spleen, the amount of Thy-1 and of immunoglobulin on cells bearing these markers was changed very little by drug treatment. The proportion of splenic B cells was diminished by treatment with cyclophosphamide and, to a lesser extent, by HCA, while the proportion of spleen cells bearing detectable Thy-1 and Lyt-1 increased correspondingly. The proportion of cells bearing Lyt-2 was altered by only two drugs; cyclophosphamide increased both Lyt-1+2+ and Lyt-1-2+ spleen cells and ABPP (an interferon inducer which stimulates antibody formation) decreased both Lyt-2+ subpopulations. Treatment with two drugs caused the serum corticosterone concentration to rise: ABPP increased serum corticosterone substantially while the prostaglandin induced a smaller and more transitory increase. An indirect mechanism, via corticosteroid release, might explain the thymic depletion observed in mice treated with 15(S)-methyl PGE1 and ABPP, but neither the suppression of the GVH reaction by these drugs nor polyclonal activation of B cells by ABPP can be attributed to endogenous corticosteroids. Our data show that enumeration of splenic lymphocyte subpopulations by immunofluorescence techniques may aid in elucidating the mode of action of immunomodulatory drugs.
Subject(s)
Immunosuppressive Agents/pharmacology , Lymphocytes/drug effects , Spleen/drug effects , Thymus Gland/drug effects , Animals , Antibody Formation/drug effects , Corticosterone/blood , Corticosterone/pharmacology , Cyclophosphamide/pharmacology , Cytarabine/pharmacology , Female , Fluorescent Antibody Technique , Graft vs Host Reaction/drug effects , Hydrocortisone/pharmacology , Lymphocytes/classification , Lymphocytes/immunology , Mice , Mice, Inbred C57BL , Mice, Inbred CBA , Pyrimidinones/pharmacology , Spleen/cytology , Spleen/immunology , Thymus Gland/cytology , Thymus Gland/immunologyABSTRACT
Female MRL-Mp-lpr/lpr mice spontaneously develop autoimmune disease at three to five months of age and die most commonly from immune complex glomerulonephritis. Kidneys of two-month-old females appeared nearly normal by electron microscopy, and glomerular deposits of IgG an complement component 3 (C3) barely were detectable. In five-month-old females, immunofluorescence revealed numerous deposits of IgG and C3; glomerular mesangial cells were hypertrophic and hyperplastic and contained electron-dense material. There were subepithelial and subendothelial deposits of electron-dense material with swelling of epithelial cell cytoplasm. This disease has many features similar to the immune complex glomerulonephritis observed in New Zealand Black and White hybrid mice and in man.
Subject(s)
Autoimmune Diseases/veterinary , Glomerulonephritis/veterinary , Mice, Inbred Strains , Rodent Diseases/pathology , Animals , Autoimmune Diseases/pathology , Female , Glomerulonephritis/pathology , Immune Complex Diseases/pathology , Immune Complex Diseases/veterinary , Kidney Glomerulus/ultrastructure , MiceABSTRACT
We have used spin labeling, fluorescence polarization, and chemical analysis to characterize membrane properties of thymocytes from mice treated with immunomodulatory drugs. The number of thymocytes was reduced 90-95% by treatment of 6-9 week old mice with hydrocortisone acetate (HCA) or methylprednisolone (both 125 mg/kg) or with cyclophosphamide (250 mg/kg). Electron spin resonance (esr) examination of thymocytes labeled with 5-nitroxyl stearic acid indicated that the membranes of cells remaining after treatment with any of these drugs were more rigid than those from saline-treated controls. The total cholesterol/phospholipid (C/PL) molar ratio of the HCA-resistant thymocytes was twice that of the control mice. Treatment of mice with other immunomodulatory drugs, cyclophosphamide, cytosine arabinoside (Ara-C), 2-amino-5-bromo-6-phenyl-4-(3H)-pyrimidinone (ABPP) and 15(S)-methyl prostaglandin E1 (15(S)-methyl PGE1), also altered the C/PL ratio in thymocytes and, in some cases, in spleen cells. Fluorescence polarization measurements of thymocytes labeled with 1,6-diphenyl-1,3,5-hexatriene (DPH) did not reveal the differences between cells from HCA-and saline-treated mice that were detected by spin labeling and chemical analysis. Our results indicate that the greater rigidity detected by spin labeling of hydrocortisone-resistant thymocytes may be due, at least in part, to greater membrane cholesterol content. Of the methods employed, chemical analysis was the most sensitive in revealing drug-induced alterations in thymocyte populations.
Subject(s)
Alprostadil/analogs & derivatives , Cholesterol/analysis , Immunosuppressive Agents/pharmacology , Spleen/drug effects , Thymus Gland/drug effects , Animals , Cell Membrane/analysis , Cell Membrane/drug effects , Cyclophosphamide/pharmacology , Cytarabine/pharmacology , Electron Spin Resonance Spectroscopy , Female , Hydrocortisone/analogs & derivatives , Hydrocortisone/pharmacology , Male , Methylprednisolone/pharmacology , Mice , Mice, Inbred AKR , Mice, Inbred C57BL , Mice, Inbred CBA , Prostaglandins E, Synthetic , Pyrimidines/pharmacology , Spectrometry, Fluorescence , Spleen/analysis , Spleen/cytology , Thymus Gland/analysis , Thymus Gland/cytologyABSTRACT
The number and functional abilities of lymphocytes from patients with either breast cancer or cervical cancer were assessed before, during and after radiation therapy given to treat the disease and the patients were followed for several years. Radiation depleted both T and B cells and depressed the responses to PHA and Con-A in all patients groups. The response to mitogens in the weeks immediately following radiation therapy was greater in patients whose disease did not recur and was depressed for a much longer time in patients whose disease recurred within the next few years. This difference was most clearly reflected in the ratio of mitogen response to the number of T-cells. The relationship of responsive to non-responsive or suppressor lymphocytes in the weeks following radiation therapy may be a clue to prognosis.