Hypersensitivity Skin Test Response Research Articles

Stool Microbiota and Vaccine Responses of Infants

Oral vaccine efficacy is low in less-developed countries, perhaps due to intestinal dysbiosis. This study determined if stool microbiota composition predicted infant oral and parenteral vaccine responses. The stool microbiota of 48 Bangladeshi infants was characterized at 6, 11, and 15 weeks of age by amplification and sequencing of the 16S ribosomal RNA gene V4 region and by Bifidobacterium-specific, quantitative polymerase chain reaction. Responses to oral polio virus (OPV), bacille Calmette-Guérin (BCG), tetanus toxoid (TT), and hepatitis B virus vaccines were measured at 15 weeks by using vaccine-specific T-cell proliferation for all vaccines, the delayed-type hypersensitivity skin-test response for BCG, and immunoglobulin G responses using the antibody in lymphocyte supernatant method for OPV, TT, and hepatitis B virus. Thymic index (TI) was measured by ultrasound. Actinobacteria (predominantly Bifidobacterium longum subspecies infantis) dominated the stool microbiota, with Proteobacteria and Bacteroidetes increasing by 15 weeks. Actinobacteria abundance was positively associated with T-cell responses to BCG, OPV, and TT; with the delayed-type hypersensitivity response; with immunoglobulin G responses; and with TI. B longum subspecies infantis correlated positively with TI and several vaccine responses. Bacterial diversity and abundance of Enterobacteriales, Pseudomonadales, and Clostridiales were associated with neutrophilia and lower vaccine responses. Bifidobacterium predominance may enhance thymic development and responses to both oral and parenteral vaccines early in infancy, whereas deviation from this pattern, resulting in greater bacterial diversity, may cause systemic inflammation (neutrophilia) and lower vaccine responses. Vaccine responsiveness may be improved by promoting intestinal bifidobacteria and minimizing dysbiosis early in infancy.

Hepatitis B vaccine antibody response and the risk of clinical AIDS or death.

BackgroundWhether seroresponse to a vaccine such as hepatitis B virus (HBV) vaccine can provide a measure of the functional immune status of HIV-infected persons is unknown.This study evaluated the relationship between HBV vaccine seroresponses and progression to clinical AIDS or death.Methods and FindingsFrom a large HIV cohort, we evaluated those who received HBV vaccine only after HIV diagnosis and had anti-HBs determination 1–12 months after the last vaccine dose. Non-response and positive response were defined as anti-HBs <10 and ≥10 IU/L, respectively. Participants were followed from date of last vaccination to clinical AIDS, death, or last visit. Univariate and multivariable risk of progression to clinical AIDS or death were evaluated with Cox regression models. A total of 795 participants vaccinated from 1986–2010 were included, of which 41% were responders. During 3,872 person-years of observation, 122 AIDS or death events occurred (53% after 1995). Twenty-two percent of non-responders experienced clinical AIDS or death compared with 5% of responders (p<0.001). Non-response to HBV vaccine was associated with a greater than 2-fold increased risk of clinical AIDS or death (HR 2.47; 95% CI, 1.38–4.43) compared with a positive response, after adjusting for CD4 count, HIV viral load, HAART use, and delayed type hypersensitivity skin test responses (an in vivo marker of cell-mediated immunity). This association remained evident among those with CD4 count ≥500 cells/mm3 (HR 3.40; 95% CI, 1.39–8.32).ConclusionsHBV vaccine responses may have utility in assessing functional immune status and risk stratificating HIV-infected individuals, including those with CD4 count ≥500 cells/mm3.

Development of a Skin Test for Bovine Tuberculosis for Differentiating Infected from Vaccinated Animals

The tuberculin skin test has been used for the diagnosis of bovine and human tuberculosis (TB) for over a hundred years. However, the specificity of the test is compromised by vaccination with the Mycobacterium bovis-derived vaccine strain bacille Calmette-Guérin (BCG). Since current promising vaccines against bovine TB are based on heterologous prime-boost combinations that include BCG, there is a need for diagnostic tests for differentiating infected from vaccinated animals (DIVA). The application of antigens such as ESAT-6 and CFP-10 for DIVA has so far been realized largely through their application in the blood-based gamma interferon release assay. In the current study, we have reassessed the potential of such antigens as skin test reagents for DIVA in cattle. A cocktail of the Mycobacterium tuberculosis complex recombinant protein antigens ESAT-6, CFP-10, MPB70, and MPB83 elicited delayed-type hypersensitivity (DTH) skin test responses in 78% of naturally infected tuberculin-positive cattle. Importantly, this cocktail induced no skin responses in BCG-vaccinated cattle despite them being sensitized for strong tuberculin responses. Further optimization of skin test antigen combinations identified that the inclusion of Rv3615c (Mb3645c) enhanced skin test sensitivity in naturally infected cattle without compromising specificity. In addition, we demonstrate for the first time the utility of synthetic peptides as promising skin test antigens for bovine TB for DIVA. Our data provide a promising basis for the future development of skin tests for DIVA with practical relevance for TB diagnosis in both veterinary and clinical settings.

Independent effects of genetic variations in mannose-binding lectin influence the course of HIV disease: the advantage of heterozygosity for coding mutations.

The in vivo impact of mannose-binding lectin (MBL), a molecule involved in innate immunity, on the pathogenesis of human immunodeficiency virus (HIV)-1 infection and AIDS is unknown. A total of 1102 HIV-positive and 2213 HIV-negative adult subjects were screened for polymorphisms in the coding and promoter regions of MBL2, the gene that encodes MBL. Variations in MBL2 did not influence the risk of acquiring HIV-1. Heterozygosity for coding mutations (O allele) and homozygosity for the -221 promoter polymorphism (X allele) in MBL2 were associated with a delay in and an accelerated rate of disease progression, respectively. MBL2 variations influenced the rate of progression to AIDS-defining illnesses. In a multivariate model, the effects of MBL2 variations were independent of several parameters known to influence disease progression, including steady-state viral load, baseline CD4(+) T cell counts, and delayed-type hypersensitivity skin test responses, an in vivo marker of cell-mediated immunity. The effects of MBL2 variations were most evident in those who possessed protective genotypes of CCR5 and a high copy number of CCL3L1, the most potent HIV-suppressive CCR5 ligand. MBL2 genotypes are independent determinants of HIV disease progression and heterozygosity for MBL2 coding mutations confer disease-retarding effects. MBL-dependent immune responses may play a role in the pathogenesis of HIV infection.

Genetic Predisposition to Self‐Curing Infection with the ProtozoanLeishmania chagasi:A Genomewide Scan

The protozoan Leishmania chagasi can cause disseminated, fatal visceral leishmaniasis (VL) or asymptomatic infection in humans. We hypothesized that host genetic factors contribute to this variable response to infection. A family study was performed in neighborhoods of endemicity for L. chagasi near Natal in northeastern Brazil. Study subjects were assessed for the presence of VL or asymptomatic infection, which was defined by a positive delayed-type hypersensitivity (DTH) skin test response to Leishmania antigen without disease symptoms. A genomewide panel of 385 autosomal microsatellite markers in 1254 subjects from 191 families was analyzed to identify regions of linkage. Regions with potential linkage to the DTH response on chromosomes 15 and 19, as well as a novel region on chromosome 9 with potential linkage to VL, were identified. Understanding the genetic factors that determine whether an individual will develop symptomatic or asymptomatic infection with L. chagasi may identify proteins essential for immune protection against this parasitic disease and reveal strategies for immunotherapy or prevention.

Genes at human chromosome 5q31.1 regulate delayed-type hypersensitivity responses associated with Leishmania chagasi infection

Visceral leishmaniasis (VL) caused by Leishmania chagasi is endemic to northeast Brazil. A positive delayed-type hypersensitivity skin test response (DTH+) is a marker for acquired resistance to disease, clusters in families and may be genetically controlled. Twenty-three single nucleotide polymorphisms (SNPs) were genotyped in the cytokine 5q23.3-q31.1 region IRF1-IL5-IL13-IL4-IL9-LECT2-TGFBI in 102 families (323 DTH+; 190 DTH-; 123 VL individuals) from a VL endemic region in northeast Brazil. Data from 20 SNPs were analyzed for association with DTH+/- status and VL using family-based, stepwise conditional logistic regression analysis. Independent associations were observed between the DTH+ phenotype and markers in separate linkage disequilibrium blocks in LECT2 (OR 2.25; P=0.005; 95% CI=1.28-3.97) and TGFBI (OR 1.94; P=0.003; 95% CI=1.24-3.03). VL child/parent trios gave no evidence of association, but the DTH- phenotype was associated with SNP rs2070874 at IL4 (OR 3.14; P=0.006; 95% CI=1.38-7.14), and SNP rs30740 between LECT2 and TGFBI (OR 3.00; P=0.042; 95% CI=1.04-8.65). These results indicate several genes in the immune response gene cluster at 5q23.3-q31.1 influence outcomes of L. chagasi infection in this region of Brazil.

Induction of Peptide-Specific Immune Response in Patients with Primary Malignant Melanoma of the Esophagus after Immunotherapy Using Dendritic Cells Pulsed with MAGE Peptides

Primary malignant melanoma of the esophagus (PMME) is a very rare disease with an extremely poor prognosis. Surgery is currently considered its best treatment, while any other measures are ineffective. We studied the effect of active specific immunotherapy using monocyte-derived dendritic cells (DCs) pulsed with the epitope peptides of melanoma-associated antigens (MAGE-1, MAGE-3) in patients with PMME after surgery, for the first time. The patient received passive immunotherapy with lymphokine-activated killer cells concomitantly. Two HLA-A24-positive patients with PMME were treated. Both patients initially received radical esophagectomy with regional lymphadenectomy, followed by adjuvant chemotherapy with dacarbazine, nimustine, vincristine and interferon-alpha. In the case 1 patient, active specific immunotherapy was used to treat a large abdominal lymph node metastasis that became obvious 21 months after surgery. The disease remained stable for 5 months, and the patient survived for 12 months after the initiation of immunotherapy. In the case 2 patient, immunotherapy was tried as post-operative adjuvant treatment after adjuvant chemotherapy. There was no tumor recurrence for 16 months after the immunotherapy. As of 49 months after esophagectomy, the patient is still alive. In both patients, the ability of peripheral lymphocytes to produce IFN-gamma in vitro in response to peptide stimulation was significantly enhanced and delayed-type hypersensitivity skin test response to MAGE-3 peptide was turned positive after immunotherapy. In conclusion, active specific immunotherapy for PMME with the use of DCs and MAGE peptides was safe and capable of inducing peptide-specific immune responses. This case report warrants further clinical evaluation of this immunotherapy for PMME.

Vitamin E and Respiratory Infection in the Elderly

Respiratory infections are prevalent in the elderly, resulting in increased morbidity, mortality, and utilization of health care services. Contributing to the increased incidence of infection with age is the well-described decline in immune response, which has been correlated with patterns of illness in the elderly. For example, there are higher morbidity and mortality from cancer, pneumonia, and post-operative complications in those who have diminished, delayed-type hypersensitivity skin test responses. Nutritional status is an important determinant of immune function. We have shown in double-blind, placebo-controlled trials that vitamin E supplementation significantly improved immune response, including DTH and response to vaccines. Furthermore, subjects receiving vitamin E in the 6-month trial had a 30% lower incidence of infectious diseases. That study, however, was not powered to demonstrate statistical significance, and the infections were self-reported. To overcome these limitations, we conducted a double-blind, placebo-controlled trial to determine the effect of one-year supplementation with 200 IU/day vitamin E on the incidence and duration of respiratory infections in 617 elderly nursing home residents. The results of this clinical trial show that vitamin E supplementation significantly reduces the incidence rate of common colds and the number of subjects who acquire a cold among elderly nursing home residents. A nonsignificant reduction in the duration of colds was also observed. Because of the high rate and more severe morbidity associated with common colds in this age group, these findings have important implications for the well being of the elderly as well as for the economic burden associated with their care.

Enhanced cellular immune response in women with PTSD related to childhood abuse.

Disturbed regulation of both the hypothalamic-pituitary-adrenal (HPA) axis and the sympathoadrenomedullary system in posttraumatic stress disorder (PTSD) suggests that immune function, which is modulated by these systems, also may be dysregulated in individuals with PTSD. Delayed-type hypersensitivity skin test responses were measured in 16 women with PTSD due to childhood sexual or physical abuse and 15 women who did not have a history of abuse, other trauma, or psychiatric disorders. HPA axis activity was assessed by examination of circadian salivary cortisol levels and a single time point measurement of plasma cortisol. Delayed-type hypersensitivity was enhanced in women with PTSD. Cortisol measures did not differ between PTSD and healthy comparison subjects. These results suggest that cell-mediated inflammatory reactions are greater in individuals with PTSD.

BCG-induced increase in interferon-gamma response to mycobacterial antigens and efficacy of BCG vaccination in Malawi and the UK: two randomised controlled studies

The efficacy of BCG vaccines against pulmonary tuberculosis varies between populations, showing no protection in Malawi but 50-80% protection in the UK. To investigate the mechanism underlying these differences, randomised controlled studies were set up to measure vaccine-induced immune responsiveness to mycobacterial antigens in both populations. 483 adolescents and young adults in Malawi and 180 adolescents in the UK were tested for interferon-gamma (IFN-gamma) response to M tuberculosis purified protein derivative (PPD) in a whole blood assay, and for delayed type hypersensitivity (DTH) skin test response to tuberculin PPD, before and 1 year after receiving BCG (Glaxo 1077) vaccination or placebo or no vaccine. The percentages of the randomised individuals who showed IFN-gamma and DTH responses were higher in Malawi than in the UK pre-vaccination-ie, 61% (331/546) versus 22% (47/213) for IFN-gamma and 46% (236/517) versus 13% (27/211) for DTH. IFN-gamma responses increased more in the UK than in Malawi, with 83% (101/122) and 78% (251/321) respectively of the vaccinated groups responding, with similar distributions in the two populations 1 year post-vaccination. The DTH response increased following vaccination in both locations, but to a greater extent in the UK than Malawi. The IFN-gamma and DTH responses were strongly associated, except among vaccinees in Malawi. The magnitude of the BCG-attributable increase in IFN-gamma responsiveness to M tuberculosis PPD, from before to 1 year post-vaccination, correlates better with the known levels of protection induced by immunisation with BCG than does the absolute value of the IFN-gamma or DTH response after vaccination. It is likely that differential sensitisation due to exposure to environmental mycobacteria is the most important determinant of the observed differences in protection by BCG between populations.

Relationships between immunity and dietary and serum antioxidants, trace metals, B vitamins, and homocysteine in elderly men and women

It is well known that nutritional status has profound effects on immunity. The compromised cellular immunity that occurs in many otherwise healthy older people includes impaired interleukin-2 (IL-2) production and anergy to skin test antigens. The latter has been associated prospectively with increased mortality in the elderly. We studied dietary and blood concentrations of vitamins A, B6, C and E, folate, β carotene, and zinc and their relationship to delayed hypersensitivity skin test responses (DHST) and serum soluble IL-2 receptor concentrations (IL-2R) in 65 healthy independently-living men and women aged 69.8 ± 7.2 years. Serum concentrations of vitamin B12 and copper and dietary lycopene were also measured. Excluded were subjects who had used vitamin or mineral supplements in the preceding 3 months, since supplementation may improve DHST responses. IL-2R concentrations were positively (p < 0.05) associated with serum concentrations of homocysteine, vitamin B6 and BMI, and negatively associated with serum β-carotene and dietary lycopene. In a multiple regression model, these 5 variables explained 52% of the variability in IL-2R. The percentage of subjects with anergy to a panel of 7 skin test antigens was 25%. DHST responses were negatively associated with T helper cell numbers and serum retinol, T helper cell numbers were positively associated with serum copper, and natural killer cell numbers were positively associated with dietary folate and vitamin B6. Serum homocysteine concentrations were negatively associated with plasma concentrations of folate, and vitamin B12, as well as positively with serum IL-2R. In a multiple regression model, these 3 concentrations explained 27% of the variability in serum homocysteine. The results suggest that IL-2 metabolism may be influenced by several dietary antioxidants and B vitamins including those that alter homocysteine concentrations.

Vaccination with a CDR2 BV6S2/6S5 peptide in adjuvant induces peptide-specific T-cell responses in patients with multiple sclerosis

Earlier studies from several groups including ours have documented that patients with multiple sclerosis (MS) have over-expression of activated T-cells from specific TCR V beta families, including BV6S2/S5 (Kotzin et al. [1991] Proc. Natl. Acad. Sci. USA 88:9161--9165; Gold et al. [1997] J. Neuroimmunol. 76:29--38). It has also been established in the rat EAE model that peptide vaccines to the over-expressed V beta 8.2 TCR can prevent MBP induced disease (Vandenbark et al. [1989] Nature 341:541--544). In the current clinical study, 10 patients were vaccinated with 300 microg of BV6S2/6S5 peptide emulsified in incomplete Freund's adjuvant (IFA) and monitored for safety and immunogenicity in a 48-week multicenter, open-label trial. The peptide vaccine was well tolerated and no serious adverse events were observed. Vaccinations induced cell-mediated immunity to the immunizing peptide in eight of 10 patients as demonstrated by lymphocyte proliferation assay (LPA) and delayed-type hypersensitivity (DTH) skin test responses. In summary, these results demonstrate that immunization with TCR BV6S2/6S5 peptide vaccine in MS patients is safe and immunogenic, and supports a larger double-blind placebo controlled trial to determine the clinical efficacy of this approach.

Cytokine profiles in immunocompetent persons infected with Mycobacterium avium complex.

To evaluate the immunologic factors that contribute to protection against Mycobacterium avium complex (MAC), cytokine production by peripheral blood mononuclear cells (PBMC) from human immunodeficiency virus-negative persons with pulmonary MAC (MAC patients) and healthy control subjects with a delayed hypersensitivity skin test response to M. avium sensitin (MAS-positive control subjects) was measured. In MAC patients, mycobacterium-stimulated PBMC produced higher concentrations of interleukin (IL)-10 but lower concentrations of interferon (IFN)-gamma, IL-12, and tumor necrosis factor (TNF)-alpha, compared with PBMC from MAS-positive control subjects. Immunolabeling for intracellular IL-10 revealed that this cytokine was produced by both monocytes and T cells. Alveolar macrophages produced TNF-alpha and IL-10 in response to MAC, which suggests that these cytokines are produced in the lungs of patients with pulmonary disease caused by this pathogen. Our findings suggest that IFN-gamma, TNF-alpha, and IL-12 contribute to protection against MAC, whereas IL-10 is immunosuppressive.

Immune Reconstitution in the HAART Era, Part 2: Implications for Practice

As discussed in Part 1 of this series, HIV infection alters the function of the immune system. These alterations include declines in CD4-positive lymphocyte counts, disturbances in the function of the CD4-positive lymphocyte, diminished delayed-type hypersensitivity (DTH) skin test responses and cell-mediated immune responses, phenotypic changes in lymphocytes, and changes in cytoxic T-lymphocyte and natural killer cell function. Although highly active antiretroviral therapy (HAART) has dramatically improved CD4-positive lymphocyte counts and quality of life, there have been unforeseen consequences of immune reconstitution. For example, cytomegalovirus (CMV) and hepatitis C flares have been observed in HAART-treated patients coinfected with HIV and CMV and hepatitis C (HCV). Part 2 of this series will examine the clinical consequences of immune reconstitution in persons with HIV infection.

Serum dioxin and immunologic response in veterans of Operation Ranch Hand.

The authors studied immune response and exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (dioxin) among veterans of Operation Ranch Hand, the US Air Force unit responsible for the aerial spraying of herbicides in Vietnam from 1962 to 1971. A comparison group of Air Force veterans who served in Southeast Asia but were not involved in spraying herbicides was included. The authors studied delayed-type hypersensitivity skin test responses to Candida albicans, mumps, Trichophyton, and a bacterial antigen made from lysed Staphylococcus aureus. Lymphocyte measurements included total lymphocyte counts; T-cell (CD3, CD4, CD5, and CD8), B-cell (CD20), and NK-cell (CD16 and CD56) subsets; and expression of the activation antigen CD25 on CD3 T cells. The authors quantitated the serum concentrations of immunoglobulin (Ig)A, IgG, and IgM; examined sera for the presence of monoclonal immunoglobulins (M proteins); and looked for a broad range of autoantibodies (rheumatoid factor, antinuclear antibody, smooth muscle autoantibody, mitochondrial autoantibody, parietal cell autoantibody, and thyroid microsomal autoantibodies). They measured the level of dioxin in 1987 or 1992, extrapolated the result to the time of service in Vietnam, and assigned each veteran to one of four exposure categories: Comparison and three Ranch Hand groups (Background, Low, or High). Overall, the authors found no evidence of a consistent relation between dioxin exposure category and immune system alteration.

Short- and long-term beta-carotene supplementation do not influence T cell-mediated immunity in healthy elderly persons

Supplementation of healthy elderly persons with beta-carotene has been considered a way to enhance immune responses. In study 1 the short-term effect of beta-carotene (90 mg/d for 3 wk) on immunity was assessed in a randomized, double-blind, placebo-controlled longitudinal comparison of healthy elderly women. In study 2 the long-term effect of beta-carotene (50 mg every other day for 10-12 y) on immunity was assessed in a randomized, double-blind, placebo-controlled longitudinal comparison of men enrolled in the Physicians' Health Study. Subjects from both studies taking active supplements had significantly greater plasma beta-carotene concentrations than did subjects taking placebo. The pre- to postintervention change in delayed-type hypersensitivity skin test responses between beta-carotene and placebo groups in the short-term study was not significantly different, nor was the response between treatment groups in the long-term study. There were no significant effects due to beta-carotene supplementation on in vitro lymphocyte proliferation, production of interleukin 2, or production of prostaglandin E2 as a result of short- or long-term beta-carotene supplementation. In addition, there were no differences in the profiles of lymphocyte subsets [total T cells (CD3+), T helper cells (CD4+), T cytotoxic-suppressor cells (CD8+), and B cells (CD19+)] due to short- or long-term beta-carotene supplementation, nor were there differences in percentages of CD16+ natural killer cells or activated lymphocytes (cells expressing interleukin 2 transferrin receptor) due to long-term beta-carotene supplementation. Consistent results from these two trials show that beta-carotene supplementation did not have an enhancing or suppressive effect on T cell-mediated immunity of healthy elderly.

Infection of human dendritic cells by measles virus induces immune suppression.

Measles is characterized by lifelong immunity and a transient immunosuppression which, in developing countries, is responsible for a high morbidity and a high mortality consecutive to secondary infections. Strickingly, the immune suppression is coincident with MV-specific immunity and continues for several weeks after apparent recovery from measles. The immune suppression is characterized by the loss of delayed-type hypersensitivity skin test responses to recall antigens, such as tuberculin, and by the inhibition of antibody production and cellular immune responses to new antigens (1).

Evaluation of humoral and cell-mediated inducible immunity to Haemophilus ducreyi in an animal model of chancroid

To study the mechanisms of inducible immunity to Haemophilus ducreyi infection in the temperature-dependent rabbit model of chancroid, we conducted passive immunization experiments and characterized the inflammatory infiltrate of chancroidal lesions. Polyclonal immunoglobulin G was purified from immune sera raised against H. ducreyi 35000 whole-cell lysate or a pilus preparation and from naive control rabbits. Rabbits were passively immunized with 24 or 48 mg of purified polyclonal immunoglobulin G intravenously, followed 24 h after infusion by homologous titered infectious challenge. Despite titratable antibody, no significant difference in infection or disease was observed. We then evaluated the immunohistology of lesions produced by homologous-strain challenge in sham-immunized rabbits and those protectively vaccinated by pilus preparation immunization. Immunohistochemical stains for CD5 and CD4 T-lymphocyte markers were performed on lesion sections 4, 10, 15, and 21 days from infection. Lesions of pilus preparation vaccinees compared with those of controls had earlier infiltration with significantly more T lymphocytes (CD5+) and with a greater proportion of CD4+ T lymphocytes at day 4 (33% +/- 55% versus 9.7% +/- 2%; P = 0.002), corroborating earlier sterilization (5.0 +/- 2 versus 13.7 +/- 0.71 days; P < 0.001) and lesion resolution. Intraepithelial challenge of pilus-vaccinated rabbits with 100 micrograms of the pilus preparation alone produced indurated lesions within 48 h with lymphoid and plasmacytoid infiltration, edema, and extravasation of erythrocytes. We conclude that passive immunization may not confer a vaccine effect in this model and that active vaccination with a pilus preparation induces a delayed-type hypersensitivity skin test response and confers protection through cell-mediated immunity seen as an amplified lymphocytic infiltrate and accelerated maturation of the T-lymphocyte response.

Changes in Cytokine Production after Measles Virus Vaccination: Predominant Production of IL-4 Suggests Induction of a Th2 Response

Immunization with live measles virus vaccine produces transient depression of delayed-type hypersensitivity (DTH) skin test responses and mitogen-induced lymphoproliferation irrespective of the serostatus of the recipient of the vaccine. To investigate this immune suppression further we studied peripheral blood mononuclear cells (PBMC) from adults before ( N = 17) and at various times after ( N = 34) immunization with measles virus vaccine. PHA-induced lymphoproliferation was decreased after vaccine and this was partly reversed by supplementation with rIL-2. There was no change in the proportion of PBMC that were CD4 + T cells, CD8 + T cells, NK cells, or B cells as analyzed by flow cytometry. Supernatant fluids were collected from PBMC after 72 hr in culture. Analysis for cytokines after vaccination showed spontaneous production of high levels of IL-4 (vaccinees 99 ± 23; controls 5.6 ± 5.6 ng/ml, P = 0.031) and TNFα (vaccinees 140 ± 45; controls 42 ± 14 pg/ml, P = 0.072) accompanied by low levels of IFN-γ (vaccinees 1.3 ± 0.6; controls 14.3 ± 10.1 U/ml), IL-1α (vaccinees 111 ± 22; controls 442 ± 107 pg/ml, P = 0.0001), and PGE 2 (vaccinees 75 ± 39; controls 300 ± 72 pg/ml, P = 0.048). Increased amounts of IL-4 were also produced after stimulation with PHA (vaccinees 140 ± 25; controls 40 ± 40 ng/ml, P = 0.013) while levels of IFN-γ and soluble IL-2 receptor were similar to controls and levels of IL-1α (vaccinees 443 ± 67; controls 792 ± 118 pg/ml, P = 0.026) remained low. Addition of rIL-2 had little effect on these cytokine levels. These data suggest that Th2 cells producing IL-4 are preferentially activated by measles vaccine and may contribute to the immunologic abnormalities associated with immunization for measles and possibly other viral infections.

Cell-Mediated Immune Responses of Adults to Vaccination, Challenge with Rickettsia Rickettsii, or both

As a part of a study to evaluate a formalin-killed Rickettsia rickettsii vaccine, lymphoproliferative (LT) and delayed-type hypersensitivity (DTH) skin test responses to killed R. rickettsii were measured as correlates of cell-mediated immunity in volunteers who were vaccinated, challenged with R. rickettsii, or both. We detected LT responses in 26 (51%) of 51 volunteers after vaccination. After challenge, six of six unvaccinated volunteers and 12 of 16 vaccinated volunteers developed Rocky Mountain spotted fever (RMSF); all 22 mounted LT responses. The vaccinated individuals developed LT responses of greater magnitude and 1-2 weeks earlier than unimmunized controls (41,049 versus 15,084 mean net counts per minute [cpm]), suggesting that vaccination primed the cellular immune system. Moreover, development of LT responses postvaccination was associated with the amelioration of RMSF, as indicated by a slightly longer mean incubation period (328 hr versus 302 hr) and a shorter illness (19 hr versus 26 hr) in LT responders than in LT nonresponders. However, the postvaccination LT response did not discriminate between vaccinated individuals who resisted challenge and those who did not. Skin tests using killed R. rickettsii as antigen, performed in volunteers 14-17 months postvaccination or 12-15 months after challenge, revealed a weak but significant reaction in 50% of those who had received vaccine only, and a moderately strong reaction in all vaccinated and unvaccinated volunteers who had been challenged with R. rickettsii. The relationships between induction of protective immunity against intracellular bacteria by killed and replicating organisms and LT and DTH responses are discussed.