Levels Of Cell-mediated Immunity Research Articles

Prophylactic vaccination with a live-attenuated herpes zoster vaccine in lung transplant candidates

Herpes zoster (HZ) is caused by the reactivation of varicella-zoster virus (VZV). Patients with lung transplants are at high risk for HZ owing to their immunocompromised status and the need for lifelong immunosuppression. In this study, patients on the waiting list for lung transplantation were vaccinated by a live-attenuated HZ vaccine (Zostavax, Merck Sharp & Dohme), and the safety and immunogenicity of this vaccine were studied. In total, 105 patients with end-stage pulmonary disease (ESPD) were enrolled (68 participants received 1 dose of Zostavax and 37 participants were enrolled as unvaccinated controls). Among them, 43 patients underwent lung transplantation and were followed up for further analysis. VZV immunoglobulin G antibody titers and VZV-specific cell-mediated immunity (CMI) on multiple time points before and after vaccination and before and after transplantation were measured. Immune response to Zostavax was higher in younger patients, highest within 3 months after vaccination, and not influenced by gender or type of ESPD. Age, cytomegalovirus serostatus, and immunity to VZV at baseline impacted the subsequent immune response to the vaccine. Short-term immunosuppressant treatment had strong effects on VZV CMI levels, which returned to a high level at 6 months after transplantation in vaccinated patients. Zostavax did not impact infection or rejection rate after transplantation. Zostavax was safe and induced a robust humoral and cellular response for patients awaiting lung transplantation regardless of the type of ESPD. Patients younger than the recommended vaccination age of over 50 years showed a strong response and could also benefit from pre-transplant immunization.

Induction of Genotype Cross-Reactive, Hepatitis C Virus-Specific, Cell-Mediated Immunity in DNA-Vaccinated Mice.

A universal hepatitis C virus (HCV) vaccine should elicit multiantigenic, multigenotypic responses, which are more likely to protect against challenge with the range of genotypes and subtypes circulating in the community. A vaccine cocktail and vaccines encoding consensus HCV sequences are attractive approaches to achieve this goal. Consequently, in a series of mouse vaccination studies, we compared the immunogenicity of a DNA vaccine encoding a consensus HCV nonstructural 5B (NS5B) protein to that of a cocktail of DNA plasmids encoding the genotype 1b (Gt1b) and Gt3a NS5B proteins. To complement this study, we assessed responses to a multiantigenic cocktail regimen by comparing a DNA vaccine cocktail encoding Gt1b and Gt3a NS3, NS4, and NS5B proteins to a single-genotype NS3/4/5B DNA vaccine. To thoroughly evaluate in vivo cytotoxic T lymphocyte (CTL) and T helper (Th) cell responses against Gt1b and Gt3a HCV peptide-pulsed target cells, we exploited a novel fluorescent-target array (FTA). FTA and enzyme-linked immunosorbent spot (ELISpot) analyses collectively indicated that the cocktail regimens elicited higher responses to Gt1b and Gt3a NS5B proteins than those with the consensus vaccine, while the multiantigenic DNA cocktail significantly increased the responses to NS3 and NS5B compared to those elicited by the single-genotype vaccines. Thus, a DNA cocktail vaccination regimen is more effective than a consensus vaccine or a monovalent vaccine at increasing the breadth of multigenotypic T cell responses, which has implications for the development of vaccines for communities where multiple HCV genotypes circulate.IMPORTANCE Despite the development of highly effective direct-acting antivirals (DAA), infections with hepatitis C virus (HCV) continue, particularly in countries where the supply of DAA is limited. Furthermore, patients who eliminate the virus as a result of DAA therapy can still be reinfected. Thus, a vaccine for HCV is urgently required, but the heterogeneity of HCV strains makes the development of a universal vaccine difficult. To address this, we developed a novel cytolytic DNA vaccine which elicits robust cell-mediated immunity (CMI) to the nonstructural (NS) proteins in vaccinated animals. We compared the immune responses against genotypes 1 and 3 that were elicited by a consensus DNA vaccine or a DNA vaccine cocktail and showed that the cocktail induced higher levels of CMI to the NS proteins of both genotypes. This study suggests that a universal HCV vaccine can most readily be achieved by use of a DNA vaccine cocktail.

Hepatitis B Virus Vaccine–Induced Cell-Mediated Immunity Correlates with Humoral Immune Response following Primary Vaccination during Infancy

Abstract Data on hepatitis B virus (HBV) vaccine–induced cell-mediated immunity (CMI) and humoral immune response during infancy is scarce. We assessed HBV vaccine–induced CMI among infants stratified as nonresponders (Ab against HBV surface Ag [anti-HBs] levels <10 IU/l), low-responders (anti-HBs levels 10–100 IU/l), and high-responders (anti-HBs levels ≥100 IU/l) following their primary vaccination against HBV. Moreover, we assessed the association between HBV vaccine–induced CMI and anti-HBs levels. Infants were immunized with HBV vaccine at ages 2, 4, and 6 mo. Hepatitis B surface Ag (HBsAg)-specific proliferation, intracellular cytokine production, and bulk cytokine secretion were assessed on PBMCs collected at 1 y and anti-HBs levels were measured at 1 and 2 y of age. Infants classified at 2 y of age as low-responders (n = 28) had lower median levels of secreted IFN-γ than high-responders (n = 29), 17.15 pg/ml versus 33.16 pg/ml, respectively, p = 0.009. Infants classified at 2 y of age as nonresponders (n = 15) had lower median levels of secreted TNF-α than high-responders (n = 29), 116.11 pg/ml versus 162.27 pg/ml, respectively, p = 0.032. There was a positive correlation between HBsAg-specific secreted IFN-γ levels at 1 y and anti-HBs levels at 1 and 2 y of age, rho = 0.269 and 0.302, respectively, (p = 0.019 and p = 0.01, respectively). There was a positive correlation between anti-HBs levels at age 1 y and the levels of secreted IL-10, rho = 0.297, p = 0.009. HBsAg-specific IFN-γ, IL-10, and TNF-α secretion correlated with HBV vaccine–induced humoral immune response. Assessment of CMI is a useful adjunct in demonstrating the persistence of HBV vaccine–induced memory immune response.

Relationship between cell-mediated immunity to Varicella-Zoster virus and aging in subjects from the community-based Shozu Herpes Zoster study.

Age-related declines in cell-mediated immunity (CMI) are associated with the incidence and severity of Herpes Zoster (HZ) infection. However, the level of Varicella-Zoster virus (VZV)-specific CMI associated with disease onset is unclear. This study aimed to examine factors associated with VZV-specific CMI, as measured by an interferon-gamma (IFN-γ) enzyme-linked immunospot (ELISPOT) assay, in a Japanese cohort. The study enrolled 365 subjects aged 60 years and over, all of whom were taking part in the Shozu Herpes Zoster (SHEZ) study and had undergone four sets of blood and intradermal reaction tests during a 3 year follow-up period. The VZV-specific immunity profile of each subject was assessed, and linear mixed effects models were constructed to analyze IFN-γ ELISPOT results in association with a combination of factors. The model that best explained the IFN-γ ELISPOT results was selected using the Akaike Information Criteria. The best-fit model consisted of age group as the only explanatory fixed-effect variable. The model showed that VZV-specific CMI, quantified as numbers of spots on the ELISPOT assay, among subjects aged 70-79 was on average 10.30 points lower than that among subjects aged 60-69. There was no statistically significant difference between subjects aged 70-79 and those aged 80-89. Age was the only factor significantly associated with the level of VZV-specific CMI, as measured by the IFN-γ ELISPOT assay. These results may represent an important step towards quantifying the relationship between VZV-specific CMI and the onset of HZ. J. Med. Virol. 89:313-317, 2017. © 2016 Wiley Periodicals, Inc.

Correlation between Radiological and Pathological Findings in Patients with Mycoplasma pneumoniae Pneumonia.

Studies focused on the pathological–radiological correlation of human Mycoplasma (M) pneumoniae pneumonia have rarely been reported. Therefore, we extensively reviewed the literature regarding pathological and radiological studies of Mycoplasma pneumonia, and compared findings between open lung biopsy specimen and computed tomography (CT). Major three correlations were summarized. (1) Peribronchial and perivascular cuffing characterized by mononuclear cells infiltration was correlated with bronchovascular bundles thickening on CT, which was the most common finding of this pneumonia. (2) Cellular bronchitis in the small airways accompanied with exudates or granulation tissue in the lumen revealed as centrilobular nodules on CT. (3) Neutrophils and exudates in the alveolar lumen radiologically demonstrated as air-space consolidation or ground-glass opacities. In M. pulmonis-infected mice model, pathologic patterns are strikingly different according to host cell-mediated immunity (CMI) levels; treatment with interleukin-2 lead to marked cellular bronchitis in the small airways and treatment with prednisolone or cyclosporin-A lead to neutrophils and exudates in the alveolar lumen. Patients with centrilobular nodules predominant radiologic pattern have a high level of CMI, measuring by tuberculin skin test. From these findings, up-regulation of host CMI could change radiological pattern to centrilobular nodules predominant, on the other hand down-regulation of host CMI would change radiological pattern to ground-glass opacity and consolidation. It was suggested the pathological features of M. pneumoniae pneumonia may be altered by the level of host CMI.

Evaluation of varicella zoster virus-specific cell-mediated immunity by using an interferon-γ enzyme-linked immunosorbent assay

BackgroundAdministration of the varicella vaccine induces both varicella-zoster virus (VZV)-specific humoral and cell-mediated immunity (CMI). ObjectiveTo assess VZV-CMI, we developed an interferon γ enzyme-linked immunosorbent assay (IFN-γ ELISA) that measures the quantity of total IFN-γ in culture supernatants of human peripheral blood mononuclear cells. Study designWe evaluated this method by comparing the pre- and post-vaccination immune response in peripheral blood mononuclear cells of 30 healthy children who were administered an initial varicella vaccination at Konan Kosei hospital. ResultsIFN-γ ELISA showed well-validated results; CMI was not detectable pre-immunization but became detectable post-immunization. Seroconversion was detected in 92.6% of subjects by the immune adherence hemagglutination test; however, half of the subjects did not display an increase in CMI levels. We also compared the incidence of breakthrough varicella and herpes zoster development between CMI post-positive and post-negative vaccinees at 1–2years after the last VZV vaccination. Eight subjects had a history of varicella or herpes zoster exposure post-VZV vaccination. Two of them with post-negative CMI contracted breakthrough varicella 15–16months after the last vaccination, even though they had sufficient VZV-specific antibody levels to be considered seropositive and seroprotected. Conversely, the others with post-positive CMI did not contract breakthrough varicella, despite experiencing extensive VZV exposure through casual contact with playmates and family. ConclusionsThe CMI data generated by this IFN-γ ELISA may accurately reflect real-world immune status, and CMI may be closely related to immunoprotection against breakthrough varicella development.

A Multiantigenic DNA Vaccine That Induces Broad Hepatitis C Virus-Specific T-Cell Responses in Mice

There are 3 to 4 million new hepatitis C virus (HCV) infections annually around the world, but no vaccine is available. Robust T-cell mediated responses are necessary for effective clearance of the virus, and DNA vaccines result in a cell-mediated bias. Adjuvants are often required for effective vaccination, but during natural lytic viral infections damage-associated molecular patterns (DAMPs) are released, which act as natural adjuvants. Hence, a vaccine that induces cell necrosis and releases DAMPs will result in cell-mediated immunity (CMI), similar to that resulting from natural lytic viral infection. We have generated a DNA vaccine with the ability to elicit strong CMI against the HCV nonstructural (NS) proteins (3, 4A, 4B, and 5B) by encoding a cytolytic protein, perforin (PRF), and the antigens on a single plasmid. We examined the efficacy of the vaccines in C57BL/6 mice, as determined by gamma interferon enzyme-linked immunosorbent spot assay, cell proliferation studies, and intracellular cytokine production. Initially, we showed that encoding the NS4A protein in a vaccine which encoded only NS3 reduced the immunogenicity of NS3, whereas including PRF increased NS3 immunogenicity. In contrast, the inclusion of NS4A increased the immunogenicity of the NS3, NS4B, andNS5B proteins, when encoded in a DNA vaccine that also encoded PRF. Finally, vaccines that also encoded PRF elicited similar levels of CMI against each protein after vaccination with DNA encoding NS3, NS4A, NS4B, and NS5B compared to mice vaccinated with DNA encoding only NS3 or NS4B/5B. Thus, we have developed a promising "multiantigen" vaccine that elicits robust CMI. Since their development, vaccines have reduced the global burden of disease. One strategy for vaccine development is to use commercially viable DNA technology, which has the potential to generate robust immune responses. Hepatitis C virus causes chronic liver infection and is a leading cause of liver cancer. To date, no vaccine is currently available, and treatment is costly and often results in side effects, limiting the number of patients who are treated. Despite recent advances in treatment, prevention remains the key to efficient control and elimination of this virus. Here, we describe a novel DNA vaccine against hepatitis C virus that is capable of inducing robust cell-mediated immune responses in mice and is a promising vaccine candidate for humans.

The Novel Replication-defective Vaccinia Virus (Tiantan Strain)–based Hepatitis C Virus Vaccine Induces Robust Immunity in Macaques

The induction of a robust neutralizing antibody (nAb) response is likely to be as essential as specific cell-mediated immunity (CMI) against multiple antigens for the development of effective preventive and therapeutic vaccines against hepatitis C virus (HCV) infection in humans. To date, no data on the immunogenicity of the replication-defective vaccinia virus (derived from the Tiantan strain) (rNTV)-based HCV vaccine in primates have been reported. This study describes in detail the immunogenicity of various vaccine candidates in rhesus macaques, including rNTV-based and replication-defective recombinant adenoviral (rAd)-based HCV vaccines, as well as HCV pseudotyped virus-like particles (HCVpp). Our data showed that rAd-HCV vaccine boosting induced robust CMI, while priming or boosting with HCVpp enhanced the antigen-specific nAb response after rAd-HCV vaccination; however, CMI was not enhanced. Vaccination includes rNTV-HCV priming induced robust antigen-specific antibody, particularly nAbs, and CMI responses. Furthermore, more robust and longer-lasting CMI and higher cytokine levels (both Th1 and Th2 types, especially IFN-γ) resulted from boosting with rAd-HCV. We conclude that the rNTV-based HCV vaccine induces robust nAbs and CMI when combined with a heterogeneous primer-booster strategy, which shows promise for development of a human HCV vaccine.

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Induction of prominent Th1 response in C57Bl/6 mice immunized with an E.coli-expressed multi T-cell epitope EgA31 antigen against Echinococcus granulosus

First step in developing an epitope-based vaccine is to predict peptide binding to the major histocompatibility complex (MHC) molecules. We performed computational analysis of unique available EgA31 sequence to locate appropriate antigenic propensity positions. T-cell epitopes with best binding affinity values of < 50% inhibitory concentration were selected using different available servers (Propred and IEDB). Peptides with 100% population coverage were selected. A DNA fragment corresponding to the furin linker enriched in Golgi apparatus was inserted sequentially between each epitope sequences in a synthetic DNA in order to cleave the chimeric protein into four separated peptides. Subsequently, the synthetic DNA was cloned into the pGEX4T-1 and pEGFP-N1 vectors and GST-ChEgA31 was expressed in E. coli strain BL21-DE3. The recombinant protein was detected by western blotting using an HRP-conjugated polyclonal anti-GST antibody. Fusion protein purified by affinity chromatography was used to raise antisera in rabbits. Results in agar gel immunodiffusion assay indicated induction of specific antibodies against multiepitope antigen in the tested rabbits. Cytokine assay was carried out in C57Bl/6 mice and the levels of cytokines were analyzed by sandwich ELISA. Interestingly, production of specific IFN-gamma was prominently higher in mice immunized with GST-ChEgA31 and pEGFP-ChEgA31 (650-1300 pg/ml) compared to control groups. No difference was observed in the level of IL-10 and IL-4 in immunized and GST control group. Challenge study with 500 live protoscolices of Echinococcus granulosus on immunized mice demonstrated protectivity level (50-60%). Based on our results, it appeared that the chimeric protein in the study was able to stimulate T-helper cell-1 (Th1) development and high level of cell mediated immunity in mice.

Varicella-zoster virus-specific cell-mediated immunity in subjects with herpes zoster

Though cell-mediated immunity (CMI) against varicella-zoster virus (VZV) is critical for prevention of the onset of herpes zoster (HZ), clinicians currently lack a simplified procedure to monitor CMI. We have recently developed an assay, called the IFN-γ release assay, and showed that it is a simple and reliable method to determine VZV-specific CMI. In the present study, we applied an IR assay to measure the VZV-specific CMI of patients with HZ. VZV-specific CMI levels were significantly high at the onset of the disease, but were decreased several weeks later. In contrast, CMI VZV-specific antibody titers increased in convalescent phase compared to those in acute phase. Thus, this technology is likely to be very useful in monitoring ongoing VZV-specific immune status.

Correlation of Cellular Immune Responses with Protection against Culture-Confirmed Influenza Virus in Young Children

The highly sensitive gamma interferon (IFN-gamma) enzyme-linked immunosorbent spot (ELISPOT) assay permits the investigation of the role of cell-mediated immunity (CMI) in the protection of young children against influenza. Preliminary studies of young children confirmed that the IFN-gamma ELISPOT assay was a more sensitive measure of influenza memory immune responses than serum antibody and that among seronegative children aged 6 to <36 months, an intranasal dose of 10(7) fluorescent focus units (FFU) of a live attenuated influenza virus vaccine (CAIV-T) elicited substantial CMI responses. A commercial inactivated influenza virus vaccine elicited CMI responses only in children with some previous exposure to related influenza viruses as determined by detectable antibody levels prevaccination. The role of CMI in actual protection against community-acquired, culture-confirmed clinical influenza by CAIV-T was investigated in a large randomized, double-blind, placebo-controlled dose-ranging efficacy trial with 2,172 children aged 6 to <36 months in the Philippines and Thailand. The estimated protection curve indicated that the majority of infants and young children with >or=100 spot-forming cells/10(6) peripheral blood mononuclear cells were protected against clinical influenza, establishing a possible target level of CMI for future influenza vaccine development. The ELISPOT assay for IFN-gamma is a sensitive and reproducible measure of CMI and memory immune responses and contributes to establishing requirements for the future development of vaccines against influenza, especially those used for children.

Better Understanding of the Immunosuppressive Link between the Lymphocytic Immune Cells and the Decreased Cell Mediated Immunity in Head and Neck Cancer Patients

The purpose of this study was to determine the phenotyping of Peripheral Blood Lymphocytes (PBL) in Head and Neck Cancers (HNCA) patients and to relate this with the level of Cell-Mediated Immunity (CMI) measured by in vitro lymphoproliferative assay, in order to evaluate immune suppression in HNCA patients and its possible mechanisms. Accordingly, one hundred twenty two HNCA patients and 100 control subjects were enrolled in this study. HNCA patients were classified into 42 nasopharyngeal carcinoma, 66 carcinoma of larynx and 14 Hypo Pharyngeal Carcinoma (HPC). For measuring CMI, Microculture Tetrazolium assay (MTT) was applied on the freshly isolated lymphocytes of HNCA patients and control group. Immunophenotyping of PBL was carried out for monitoring the blood level of CD3+, CD4+, CD8+, CD21+ cells in HNCA patients in comparison with controls. The results of both assays have been integrated, revealed the presence of remarked immune suppression in HNCA patients in comparison with the controls, especially for NasoPharyngeal Carcinoma (NPC) patients who were immunosuppressed more than other studied HNCA types. Surprisingly, NPC group showed the lowest CMI level along with the highest level of PBL subsets, particularly NPC patients expressed the highest level of CD8+ cells among HNCA. It was inferred that CD8+ cells were more likely immune suppressor rather than cytotoxic cells and this is the principal factor for inducing sustained immunosuppression in HNCA and in NPC in particular. Furthermore CD4/CD8 ratio proved to be a reliable index for assessing the immunological status of HNCA patients and more dependable index than other immunity-evaluating factors.

DNA-MVA vaccine protection after X4 SHIV challenge in macaques correlates with day-of-challenge antiviral CD4+ cell-mediated immunity levels and postchallenge preservation of CD4+ T cell memory.

The ability of vaccines to induce immunity both in mucosal and systemic compartments may be required for prevention of HIV infection and AIDS. We compared DNA-MVA vaccination regimens adjuvanted by IL-12 DNA, administered intramuscularly and nasally or only nasally. Most of the vaccinated Rhesus macaques developed mucosal and systemic humoral and cell-mediated SHIV-specific immune responses. Stimulation of mucosal anti-Env IgA responses was limited. After rectal challenge with SHIV 89.6P, all vaccinated and naive animals became infected. However, most of the vaccinated animals showed significant control of viremia and protection from CD4(+) T cell loss and AIDS progression compared to the control animals. The levels of CD4(+) and CD8(+) T cell virus-specific responses measured on the day of challenge correlated with the level of viremia control observed later during the chronic infection. Postchallenge viremia levels inversely correlated with the preservation of SHIV-specific CD4(+)/IL-2(+) and CD8(+)/TNF-alpha(+) T cells but not with CD4(+)/IFN-gamma(+) T cells measured over time after challenge. We also found that during the early chronic infection SHIV vaccination permitted a more significant preservation of both naive and memory CD4(+) T cells compared to controls. In addition, we observed a more significant and prolonged preservation of memory CD4(+) T cells after SHIV vaccination and challenge than that observed after SIV vaccination and challenge. As the antiviral immunity stimulated by vaccination is present in the memory CD4(+) T cell subpopulations, its more limited targeting by SHIV compared to SIV may explain the better control of X4 tropic SHIV than R5 tropic SIVs by vaccination.

Immunity to Pertussis 5 Years after Booster Immunization during Adolescence

We conducted a 5-year follow-up study on the persistence of pertussis-specific antibody and cell-mediated immunity after booster immunization of adolescents aged 11-13 years with a tricomponent acellular pertussis vaccine (Boostrix; trials diphtheria-tetanus-acellular pertussis [Tdap]-004/030). Cellular and humoral immunity to pertussis toxin (PT), filamentous hemagglutinin, and pertactin were measured in adolescents (age, 16 years) 5 years after booster immunization. Similar investigations were performed for control adolescents who had received only diphtheria and tetanus booster vaccination. Five years after pertussis booster vaccination, the geometric mean concentrations of immunoglobulin G (IgG) elicited by each of the 3 pertussis vaccine antigens decreased from 1-month and 3-year postvaccination levels, but with the exception of PT IgG, were still higher than the prevaccination levels. PT IgG levels were undetectable in 28% of the subjects, but 44% of those subjects still tested positive for cell-mediated immunity to PT. Filamentous hemagglutinin IgG and pertactin IgG levels were significantly higher in Tdap-boosted adolescents than in the control subjects. Antibody concentrations at 1 month after vaccination strongly predicted antibody persistence. Cell-mediated immunity levels to PT, filamentous hemagglutinin, and pertactin persisted above the prebooster levels measured 5 years earlier. The results of the present study of adolescents indicate that the interval between acellular pertussis booster immunizations might be extended beyond 5 years.

Augmenting Immune Responses to Varicella Zoster Virus in Older Adults: A Randomized, Controlled Trial of Tai Chi

To evaluate the effects of a behavioral intervention, Tai Chi, on resting and vaccine-stimulated levels of cell-mediated immunity (CMI) to varicella zoster virus (VZV) and on health functioning in older adults. A prospective, randomized, controlled trial with allocation to two arms (Tai Chi and health education) for 25 weeks. After 16 weeks of intervention, subjects were vaccinated with VARIVAX, the live attenuated Oka/Merck VZV vaccine licensed to prevent varicella. Two urban U.S. communities between 2001 and 2005. A total of 112 healthy older adults aged 59 to 86. The primary endpoint was a quantitative measure of VZV-CMI. Secondary outcomes were scores on the Medical Outcomes Study 36-item Short-Form Health Survey (SF-36). The Tai Chi group showed higher levels of VZV-CMI than the health education group (P<.05), with a significant rate of increase (P<.001) that was nearly twice that found in the health education group. Tai Chi alone induced an increase in VZV-CMI that was comparable in magnitude with that induced by varicella vaccine, and the two were additive; Tai Chi, together with vaccine, produced a substantially higher level of VZV-CMI than vaccine alone. The Tai Chi group also showed significant improvements in SF-36 scores for physical functioning, bodily pain, vitality, and mental health (P<.05). Tai Chi augments resting levels of VZV-specific CMI and boosts VZV-CMI of the varicella vaccine.

Cell mediated immunity (CMI) and post transplant viral infections — Role of a functional immune assay to titrate immunosuppression

Cell mediated immunity (CMI) was assessed by the ImmuKnow assay in 12 patients after kidney transplantation, who presented with viral infection. Treatment included lowering of immunosuppression in all cases and antiviral treatment if indicated. The assay was repeated during the follow up. The ImmuKnow assay at time of presentation of viral infections was 56.8 ± 58.2 (range 3–178; median 22) ATP ng/ml. With the clearance of viral infection and lowering of immunosuppression, the assay showed an increase in the level of CMI at 194.5 ± 118.9 (range 53–409; median 150) ATP ng/ml. There was viral clearance or stabilization in all cases and there was no incidence of allograft rejection. The ImmuKnow assay of CMI can be used to titrate initial immunosuppression reduction and its subsequent increase, in patients with viral infection after transplantation.

FC‐3 Immunity to bovine trichophytosis

Bovine trichophytosis is a mycotic cutaneous disease caused mainly by the dermatophyte Trichophyton verrucosum. Cell‐mediated immunity (CMI) is reported to play a crucial role in defense against the disease. The objective of this study was to investigate the immune response developed after administration of a vaccine containing a live immunogenic culture of T. verrucosum. For the detection of CMI, we used an antigen‐specific lymphocyte proliferation test measured by both 3H‐thymidine incorporation and flow cytometry, and specific antigen‐induced production of IFNγ. We also tested specific antibody production by an indirect ELISA test. After vaccination by either the subcutaneous, intramuscular, or intradermal route, animals developed solid in vitro CMI 1 month after revaccination. CD4+ lymphocytes were the basic cell population activated by Trichophyton antigen in vitro. IgM and IgG antibody responses were detected from postvaccination days 3 and 7, respectively. Animals expressing higher levels of CMI displayed higher ratios of IgG2:IgG1 (a Th1 pattern of immune response). Using the challenge test, protection was higher after intramuscular &gt; subcutaneous &gt;intradermal routes of administration. We conclude that administration of a vaccine containing a live immunogenic culture of T. verrucosum leads to the development of protective immunity against bovine trichophytosis, and that the results of our laboratory tests correlated well with the results of challenge tests. Funding: Ministry of Agriculture of the Czech Republic.

Cell mediated immunity and alcohol intake in Antarctic wintering personnel

The relationship between cell-mediated immunity and alcohol intake was studied in an Antarctic wintering group. Cell-mediated immune responses have previously been shown to be significantly lowered on the Antarctic Continent. The cause for this is yet to be elucidated. The cutaneous CMI Multitest was used to assess cell-mediated immunity. Carbohydrate deficient transferrin (CDT) was used to assess chronic alcohol consumption and acute consumption was self-reported. Results showed a significantly elevated incidence of anergy when compared with previous results (53 v. 6.5%). The incidence of hypoergy was similar to previously reported levels (34%). There was no relationship demonstrated between both acute or chronic alcohol consumption and levels of cell-mediated immunity using the Spearman Rank Correlation Coefficient. There was also no correlation found between CDT and self-reported alcohol intake. It is concluded that factors other than alcohol are responsible for the decrease in cell-mediated immunity in Antarctic wintering personnel.

Differential protective effect of bacillus Calmette–Guerin vaccine against multibacillary and paucibacillary leprosy in Nagpur, India

For this paper we conducted a secondary data analysis to test the hypothesis that a linear trend exists in the protective effect of bacillus Calmette–Guerin (BCG) vaccine against types of leprosy. We used data from two previous case-control studies to perform an unmatched test for linear trend. We observed that both the studies revealed a significant linear trend ( P<0.00001). One study that estimated an insignificant protective effect of BCG against paucibacillary leprosy showed a significant departure from linearity. We conclude that, the protective effect of BCG vaccination is differential across severity of leprosy as it brings about a shift in the immune response to a higher level of cell mediated immunity. We recommend that future studies dealing with the protective effect of BCG against leprosy should also conduct an analysis for trend.