Immunological Responses In Patients Research Articles

Induction of cytotoxic T-lymphocyte responses in vivo after vaccinations with peptide-pulsed dendritic cells

Vaccination of patients with cancer using dendritic cells (DCs) was shown to be effective for B-cell lymphoma and malignant melanoma. Here we provide evidence that patients with advanced breast and ovarian cancer can be efficiently vaccinated with autologous DCs pulsed with HER-2/neu– or MUC1-derived peptides. Ten patients were included in this pilot study. The DC vaccinations were well tolerated with no side effects. In 5 of 10 patients, peptide-specific cytotoxic T lymphocytes (CTLs) could be detected in the peripheral blood using both intracellular IFN-γ staining and 51Cr-release assays. The major CTL response in vivo was induced with the HER-2/neu–derived E75 and the MUC1-derived M1.2 peptide, which lasted for more than 6 months, suggesting that these peptides might be immunodominant. In addition, in one patient vaccinated with the MUC1-derived peptides, CEA- and MAGE-3 peptide-specific T-cell responses were detected after several vaccinations. In a second patient immunized with the HER-2/neu peptides, MUC1-specific T lymphocytes were induced after 7 immunizations, suggesting that antigen spreading in vivo might occur after successful immunization with a single tumor antigen. Our results show that vaccination of DCs pulsed with a single tumor antigen may induce immunologic responses in patients with breast and ovarian cancer. This study may be relevant to the design of future clinical trials of other peptide-based vaccines.

Induction of cytotoxic T-lymphocyte responses in vivo after vaccinations with peptide-pulsed dendritic cells

AbstractVaccination of patients with cancer using dendritic cells (DCs) was shown to be effective for B-cell lymphoma and malignant melanoma. Here we provide evidence that patients with advanced breast and ovarian cancer can be efficiently vaccinated with autologous DCs pulsed with HER-2/neu– or MUC1-derived peptides. Ten patients were included in this pilot study. The DC vaccinations were well tolerated with no side effects. In 5 of 10 patients, peptide-specific cytotoxic T lymphocytes (CTLs) could be detected in the peripheral blood using both intracellular IFN-γ staining and 51Cr-release assays. The major CTL response in vivo was induced with the HER-2/neu–derived E75 and the MUC1-derived M1.2 peptide, which lasted for more than 6 months, suggesting that these peptides might be immunodominant. In addition, in one patient vaccinated with the MUC1-derived peptides, CEA- and MAGE-3 peptide-specific T-cell responses were detected after several vaccinations. In a second patient immunized with the HER-2/neu peptides, MUC1-specific T lymphocytes were induced after 7 immunizations, suggesting that antigen spreading in vivo might occur after successful immunization with a single tumor antigen. Our results show that vaccination of DCs pulsed with a single tumor antigen may induce immunologic responses in patients with breast and ovarian cancer. This study may be relevant to the design of future clinical trials of other peptide-based vaccines.

CYTOKINE VARIATIONS IN PATIENTS WITH HORMONE TREATED PROSTATE CANCER

Purpose: We evaluated the immunological response in patients with hormone sensitive and refractory prostate cancer, and untreated benign prostatic hyperplasia (BPH).Materials and Methods: Serum levels of pro-inflammatory and anti-inflammatory cytokines were measured by enzyme-linked immunosorbent assay in 3 groups of patients. The groups included 18 men with a mean age of 79 years who had hormone sensitive prostate cancer, mean prostate specific antigen (PSA) plus or minus standard deviation 1.03 ± 2.65 ng./ml. and a mean of 35 months of treatment, 10 with a mean age of 86 years who had hormone refractory prostate cancer, mean PSA 27.52 ± 42.23 ng./ml. and a mean of 42 months of treatment, and 19 with a mean age of 73 years who had BPH and mean PSA 3.37 ± 2.47 ng./ml. Results were compared with those in 10 age matched, disease-free controls. In the hormone sensitive group PSA regressed to normal and there was clinical evidence of a response to hormone ablation therapy, including orchiectomy, luteinizing h...

IMMUNE RESPONSE IN PATIENTS WITH PERSISTENT CANDIDURIA AND OCCULT CANDIDEMIA

We evaluated the immunological response in patients with persistent candiduria with or without occult candidemia. Levels of Thl (pro-inflammatory interleukin [IL]-1, IL-2 and tumor necrosis factor-alpha) and Th2 (anti-inflammatory IL-4 and IL-10) cytokines were measured in the sera of patients with persistent candiduria. Polymerase chain reaction assessment of the 158 base pair candidal actin gene was used to detect Candida albicans in blood to identify occult candidemia. During a 14-month period 66 hospitalized patients with a mean age of 63 years (range 44 to 80) with persistent candiduria were evaluated. Occult candidemia developed in 27 patients (41%) as evidenced by detection of candidal actin gene in the sera by polymerase chain reaction. Risk factors included antibiotics in 27 patients (100%), central venous catheter in 22 (81%), urinary catheter in 21 (78%), total parenteral nutrition in 18 (66%), diabetes mellitus in 16 (59%) and abdominal surgery in 14 (52%). A total of 17 age matched patients with a mean age of 59 years hospitalized for elective general or vascular surgical procedures with no clinical or laboratory evidence of urinary or hematogenous fungal or bacterial infection served as controls. Serum levels of Th2 cytokines were elevated in 18 of 39 patients with persistent candiduria alone, and in 22 of 27 patients with candiduria and occult candidemia compared to controls (p<0.002). Th1 cytokines were within normal limits or slightly decreased in all patients with persistent candiduria with or without candidemia. These observations indicate that an abnormal immune response develops in patients with persistent candiduria with or without candidemia.

Inactivated Hepatitis A Vaccine (HM175 Strain)

Inactivated Hepatitis A Vaccine (HM175 Strain)

Effects of interferon-alpha treatment on hepatitis B virus antigen-specific immunologic responses in patients with chronic hepatitis B.

Studies were undertaken to evaluate the relationship between the immune responses and the effectiveness of interferon-alpha treatment in 21 patients with HBeAg-positive chronic active hepatitis. Peripheral blood mononuclear cells (PBMC), obtained on four occasions during an 8-week course of IFN-alpha therapy, were cultured with recombinant HBcAg, purified HBeAg or pokeweed mitogen (PWM). During follow-up for 6 months after therapy, clearance of serum HBeAg was observed in eight patients designated as responders. Immunological responses of PBMC obtained before treatment did not differ between responders and non-responders. In responders, IFN-gamma and anti-HBc production was depressed during therapy, but recovered to above the pretreatment level at the end of and/or after cessation of therapy, while lymphocyte proliferation was enhanced during therapy with a subsequent decline to baseline value. In non-responders, such changes were modest throughout the study, and anti-HBc response remained decreased even after cessation of therapy. These results indicate that PBMC of responders have immunologically different responses to IFN-alpha therapy when compared with non-responders.

Legionelose associada a pneumopatias em São Paulo: estudo da comprovação etiológica por isolamento e sorologia

The role of Legionella sp as an important pathogen, although reported in many countries, had not been well documented in Brazil. The main objective of the present study is to detect this organism or it's immunological response in patients with pulmonary infections in the city of São Paulo. For this purpose, specific laboratory methodology was introduced to cultivate the agent and demonstrate specific antibodies by serology. Patients from two University centers in São Paulo were studied: 100 from the Hospital Universitario, University of São Paulo with general pulmonary infections and 100 from Hospital das Clínicas, Faculdade de Medicina, University of São Paulo. The latter were selected to exclude pulmonary infections of other bacterial aetiology, and 30 of these were selected from the Renal Transplant Unit. Clinical specimens for cultures included sputum tracheal secretion, pleural, fluid, bronchial washing or lung tissue biopsy. Isolation of Legionella sp was attempted in BCYE medium with and without antibiotics, presumptive diagnosis by biochemical methods and identification through direct immunofluorescence staining and slide agglutination test. Direct evidence of the organism in tissue was attempted by direct immunofluorescence staining. Specific antibodies for Legionella pneumophila serogroup 1 were tested by the indirect immunofluorescence assay. Serology was also carried out in specimens from human contacts with Legionnaires' Disease. Legionella pneumophila serogroup 1 was isolated from two patients, demonstrating the presence of the pathogen in this country. Serology was able to establish present or past infection with the agent in 16 of the 100 patients from Hospital das Clínicas and in only one from Hospital Universitário. In patients considered as high risk groups from Hospital das Clínicas with renal transplantation, serology for present or past Legionellosis was positive in 33% and in 8.5% for community acquired infections. Serology in specimens from Medical personnel in contact with patients of Legionnaires' disease was positive for past infection in one single subject, confirming information already published that direct transmission from person to person is unusual. The introduction of specific methods for laboratorial evidence of Legionella sp infections at the Hospital das Clínicas-Faculdade de Medicina USP community has permitted diagnosis in able time to allow use of specific anti-microbial therapy, with good results.

Immunological responses in patients with tuberculosis and in vivo effects of acetyl‐L‐carnitine oral administration

Tuberculosis (TBC) is characterized by a complex immune response which parallels the clinical course of the disease. In this respect, acquired resistance, delayed hypersensitivity reaction and anergy are the main types of immune reactivity to mycobacterial antigens. In view of the presence of nonspecific and specific immune deficits in TBC patients, a clinical trial was carried out in a group of 20 individuals with active pulmonary TBC by oral administration of acetyl-L-carnitine (ALC). This drug, which has been shown to possess immunomodulating activities, was able to upregulate the T-dependent antibacterial activity in TBC patients after 30 days' treatment, while the same activity decreased in patients receiving placebo only. On the other hand, ALC did not modify serum levels of tumour necrosis factor-α, in the same individuals. This cytokine plays a detrimental rather than beneficial role in TBC pathogenesis. In the light of these data, ALC seems to be a powerful immunomodulator in the course of Mycobacterium tuberculosis infection and other mycobacteriosis.

Partial suppression of cell mediated immunity in chromoblastomycosis.

The cellular immune response of 8 patients from the Brazilian Amazon region with chromoblastomycosis was analyzed. Primary immunological responses of patients were tested by contact sensitization to 2,4-dinitro-chlorobenzene (DNCB), or rejection of first set skin allografts. 2 of 8 patients were reactive to DNCB after sensitization, and skin allograft rejection occurred in an average of 14 days. Capacity of patients to mount recall immunological responses was measured by skin testing with two fungal antigens and three bacterial antigens. Delayed skin reaction to trichophytin and candida antigens was negative in the majority of the patients. However, reactivity to mycobacterial (tuberculin), and bacterial (staphylococcal, streptococcal) antigen was high, or only slightly diminished respectively. The data suggest that patients with chromoblastomycosis have suppressed nonspecific, cell mediated immunity for some antigens (skin allografts, DNCB, fungal antigens), while reactivity to bacterial and mycobacterial antigens is not impaired.

Urinary cytokines following photodynamic therapy for bladder cancer a preliminary report

This preliminary study was undertaken to test for the presence of urinary cytokines whose detection would provide evidence in support of the theory that photodynamic therapy (PDT) produces an immunologic response in patients treated for bladder cancer. Gamma interferon, interleukin 1-beta, interleukin 2, and tumor necrosis factor-alpha were assayed for in the urine of 4 patients treated with photodynamic therapy for bladder cancer, in 7 control patients undergoing transurethral surgical procedures, and in 5 healthy control subjects. Quantifiable concentrations of all cytokines, except gamma interferon, were measured in urine samples from the PDT patients with the highest light energies, while no urinary cytokines were found in the PDT patient who received the lowest light energy nor in any of the control subjects. These findings suggest that a local immunologic response may occur following PDT for bladder cancer.

Differential immunological response of patients with rheumatoid arthritis towards two different Epstein-Barr virus strains: inhibition of interleukin-1 release by the B95-8, but not the P3HR-1 virus strain

An abnormal immune response towards the Epstein-Barr virus (EBV) has been documented in patients with rheumatoid arthritis (RA). To investigate whether these findings are due to the transformation event caused by EBV, RA blood mononuclear cells and monocyte-enriched preparations were incubated with two different EBV strains: the transforming virus secreted by the cell line B95-8 and the virus released by the P3HR-1 cell line that is not able to transform due to a small deletion in the U2 region of the virus genome. Immunological response was determined by the production of interleukin-1 (IL-1) and tumor necrosis factor (TNF) using ELISA and bioassay systems. There was a striking difference in cytokine measurements with a strong inhibition of IL-1 and TNF production after incubation with the B95-8 virus, but not the P3HR-1 virus. These data indicate that the disturbed reaction of the immune system towards EBV is either dependent on the full transformation of B cells in RA patients or alternatively due to the secretion of a cytokine inhibitor by the B95-8 cell line.

Partial characterization of a rapidly released antigenic/allergenic component (Ag 5) of Aspergillus fumigatus

An antigenic component of Aspergillus fumigatus, which was responsible for inducing an initial early antibody response in rabbits immunized with A. fumigatus germlings, has been identified and partially characterized. By immunofluorescent antibody and ELISA techniques, this antigen was demonstrated to be present on the germling surface, although it was also detected in supernatants of conidia/germlings within 1 hour and was present in all shake and stationary culture-filtrate extracts. By incorporation of a monospecific antiserum to this component in an intermediate gel of the reference self-crossed radioimmunoelectrophoresis pattern for allergic bronchopulmonary aspergillosis sera, it was recognized as Ag 5 and was also demonstrated to bind specific IgE. Further immunochemical analyses have revealed that Ag 5 is relatively heat labile, does not bind to concanavalin A, and has a molecular weight of approximately 35 kd. This rapidly released antigenic/allergenic component may play an important role in the initiation of immunologic responses in patients with allergic bronchopulmonary aspergillosis.

Schizophrenia and multiple sclerosis.

Similarities in clinical course, age of onset, geographical distribution, and immunological responses of patients with schizophrenia and multiple sclerosis (MS) suggest that these two common illnesses of young adults may belong to a similar class of disorders. This review examines some of the similarities and differences between these two disorders and suggests that epidemiological, immunological, and viral studies considered useful in the investigation of causal factors in multiple sclerosis may also be pertinent to the search for causes of schizophrenia.

Enzyme profile and immunochemical characterization of Aspergillus fumigatus antigens

We have compared the immunochemical characteristics of culture-filtrate antigens (Ag) from Aspergillus fumigatus extracted in our laboratory with commercially available Ags. A total of 20 different preparations were studied for protein and carbohydrate content, presence of endotoxins, mycotoxins, and hemolytic toxins. These extracts were analyzed by two-dimensional electrophoresis for protein components. The immunogenicity of the preparations was determined by rocket electrophoresis with rabbit anti- A. fumigatus sera and by agar gel diffusion with sera from patients with allergic bronchopulmonary aspergillosis, aspergilloma, and normal control subjects. In order to have dependable immunologic results, the Ags must be sufficiently pure and reproducible. Until such time as pure and standardized Ags are available, the crude Ags used should be characterized to the extent that adequate reproducibility between preparations can be ascertained. The enzyme profile of the Ag preparations provides a fair indication of the quality of antigenic components, and together with other immunochemical parameters, it will be of use in determining the suitability of the extracts in immunodiagnosis. Immunochemical results demonstrate that commercial Ags contain less proteins and carbohyrates and fewer enzymes than the homemade antigens. In addition, fewer patients demonstrated specific precipitins against commercial Ags than with homemade Ags. This study once again confirms the need for pure standardized Ags for studying the immunologic response in patients with Aspergillus-induced diseases. Until such preparations are readily available, partially purified or crude Ags with known immunochemical properties and enzyme profile may be the choice for immunodiagnosis.

Immunologic response of patients with legionellosis against major protein-containing antigens of Legionella pneumophila serogroup 1 as shown by immunoblot analysis.

Major protein-containing antigens of Legionella pneumophila serogroup 1 were were identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblot analysis with rabbit antisera to 14 different Legionella species or serogroups. Fourteen bands were observed in immunoelectropherograms of whole-cell, sonicated cell, and heated cell preparations, seven of which appeared in the supernatant fluid from the heated cells and three of which were shown in an outer membrane fraction. Immunoblots of whole-cell antigen preparations of 14 Legionella species or serogroups revealed seven major Legionella proteins: antigens with molecular weights of 58,000, 79,000, and 154,000 were present in all Legionella sp. strains, antigens with molecular weights of 44,000 and 97,000 occurred in multiple species, and antigens with molecular weights of 14,000 and 25,000 were present only in L. pneumophila strains. All sera from 15 patients with culture-confirmed L. pneumophila serogroup 1 disease and 14 of 18 (78%) sera from serologically diagnosed patients reacted with the 58-kilodalton (kDa) common antigen. In contrast, less than one-half of the sera reacted with the L. pneumophila-specific proteins (14 and 25 kDa). Absorption of sera with Escherichia coli cells had no effect on their reactivity with the 58-kDa antigen, whereas absorption with L. pneumophila serogroup 1 cells removed reactivity. These data suggest that the 58-kDa antigen may prove useful in serodiagnostic tests for legionellosis.

Evaluation of blocking mechanisms against immunological responses in patients with laryngeal carcinoma.

The specific tumor-induced leukocyte inhibition factor (LIF) production in laryngeal cancer patients has been investigated before and after the removal of adherent cells in order to evaluate the existence of a suppressor activity; 20 patients served as subject. The LIF production, after challenging the lymphocytes with 3MKC1 autologous tumor extracts, was significant in 12 patients and showed a further significant increase after the removal of adherent cells. In 3 patients with no previous significant LIF production, there was a conversion to significance when the adherent cells were removed. The other patients did not show any significant variation. These data seem to suggest the existence of a suppressor activity exerted by adherent cells in laryngeal cancer patients on LIF production.

Immunological Response in Patients with Head and Neck Cancer

Immunological Response in Patients with Head and Neck Cancer

Viral oncolysate in the management of malignant melanoma.I. Preparation of the oncolysate and measurement of immunologic responses

Primary explants of human malignant melanoma were utilized in the preparation of oncolysates by Newcastle disease virus. The concentrated lysate, administered parenterally, was employed in an effort to augment antitumor immunologic responses in patients with metastatic melanoma. Observed cellular changes suggested a benefit, but humoral antibody measurements were not impressive.

Immunologic responses in patients with pulmonary carcinoma receiving immunotherapy

This is a preliminary report on immune responses observed in vivo and in vitro in resectable patients treated with oral BCG, and in nonresectable patients treated with autologous tumour cells complexed with PPD and with BCG applied by scarification. In some cases, to shorten the period of sensitization, transfer factor was given additionally. Several of the in vitro methods used for assessing the immune competence of the patients are shown to be sufficiently informative to warrant their use before immunotherapy in order to assess the immunologic resources of the patient. During immunotherapy, they allow monitoring the effect of the treatment.

Hyperthermia and Cellular Immunity

To the Editor.— In addition to conventional therapeusis employed for the treatment of cancer (eg, chemotherapy, radiation therapy, and surgery), the use of hyperthermia and hypothermia as a means of selectively destroying malignant cells received increased attention in the 1960s. 1 Renewed interest in hyperthermia isexemplified by the recent article of LeVeen et al (235:2198, 1976) and an editorial inThe Journal(235:2223, 1976). Earlier studies by Strauss et al 2 suggested induction of an immunologic response in patients with rectal carcinoma after fulguration of their tumor as one possible explanation for observations of regression of tumor in patients so treated. As one of several approaches in evaluating the effect of hyperthermia on immunologic responsiveness, we have compared the reactivity of leukocytes from patients with genitourinary tumors to unheated and heated allogeneic tissue extracts. Employing antigen-induced leukocyte adherence inhibition 3 with 3M potassium chloride extracts of allogeneic carcinomatous tissue,