Detection and characterization of IgE-producing cells in patients with clonorchiasis

The ability of peripheral B cells of patients with Clonorchis sinensis infections to secrete IgE spontaneously was investigated in vitro. The de novo synthesis of IgE was observed in unstimulated B-cell cultures of patients. There was a significant relationship between the serum IgE level and the amount of IgE spontaneously secreted by B cells. Pretreatment of patients' B cells with 10 micrograms/ml of rabbit anti-human IgE resulted in the clear suppression of spontaneous IgE synthesis without affecting the IgG synthesis. Their B cells capable of spontaneously secreting IgE were partially sensitive to irradiation with 1,000 rad. The results obtained suggest that such IgE-forming cells may be responsible for at least part of the persistent IgE formation in patients with helminthic infections as well as in those with atopic disease.     

Characterization of atypical lymphocytes and immunophenotypes of lymphocytes in patients with dengue virus infection

To characterize the immunophenotypes of lymphocytes in patients with dengue infection, we performed flow cytometric analysis of peripheral blood mononuclear cells collected from 49 dengue hemorrhagic fever (DHF), 25 dengue fever (DF), and 26 dengue-like syndrome (DLS) cases. The mean total atypical lymphocytes in DHF (916.1 +/- 685.6 cells/microl) and DF (876.2 +/- 801.9 cells/microl) were higher than those of DLS (310.5 +/- 181.4 cells/microl). An atypical lymphocyte count of 10% or higher was a good indicator of dengue infection (sensitivity 50% and specificity 86%). Flow cytometric studies showed that the percentages of atypical lymphocytes correlated with those of CD19+ B lymphocytes and inversely correlated with the percentages of CD69+ lymphocytes. The mean absolute counts of atypical lymphocytes and CD19+ cells on the discharge day were significantly higher than those on the admission day. Low percentages of TdT+ cells were found in all groups of patients. We concluded that atypical lymphocyte and CD19+ cell counts may be a useful diagnostic tool for dengue infection and the recovery from the disease could be judged when numbers of both cell types are significantly elevated.     

Phenotype analysis of tumour-infiltrating lymphocytes and lymphocytes in peripheral blood in patients with renal carcinoma

INTRODUCTION: When checking tumour growth, a number of observations indicate that the immune system plays a significant role in patients with renal cell carcinoma (RCC). Infiltration by lymphocytes (tumour infiltrating lymphocytes, TILs) is more prevalent in RCC than any other tumours. T lymphocytes are the dominant population of TIL cells. Views concerning the role of T lymphocytic subpopulations, B lymphocytes and NK cells in an anti-tumour response are not established. AIM: The aim is to determine the phenotype and activation of T and B lymphocytic subpopulations and NK cells and to compare their representation in tumour stroma and peripheral blood lymphocytes (PBL) in patients with RCC. MATERIAL AND METHODS: Samples of peripheral blood taken from the cubital and renal veins and tumour stroma cells were obtained from 44 patients in the course of their surgeries carried out due to primary RCC. TILs were isolated from mechanically disintegrated tumour tissue. Immunophenotype multiparametric analysis of PBL and TILs was carried out. Their surface and activation characteristics were determined by means of flow cytometer. RESULTS: CD3+ T lymphocytes (69.7%) were the main population of TILs. The number of CD3+/CD8+ T lymphocytes was significantly higher in TILs, 42.6% (p < 0.01), while CD4+ T lymphocytes were the majority population in peripheral blood, 41.35% (p < 0.001). The representation of CD3+/69+ T lymphocytes was significantly higher in TILs, 32.9%, compared to PBL (p < 0.001). On the contrary, the numbers of CD3+/CD25+, CD8+/57+ and CD4+/RA+ (naive CD4+ T lymphocytes) were higher in PBL (p < 0.001). The differences in representation of (CD3-/16+56+) NK cells and CD3+/DR+ T cells in TILs and PBL were not significant. CONCLUSION: The above-mentioned results prove that the characteristics and intensity of anti-tumour responses are different in compared compartments (tumour/PBL). CD3+/CD8+ T lymphocytes are the dominant lymphocytic population of TILs. The knowledge of the phenotype and functions of effector cells, which are responsible for anti-tumour response, are the basic precondition for understanding the anti-tumour immune response and the cause of its failure.     

Purine metabolism in lymphocytes from patients with primary hypogammaglobulinaemia

The previous report of low levels of purine 5'-nucleotidase activity in peripheral blood mononuclear cells (lymphocytes and monocytes) from patients with non-familial adult onset `variable' primary hypogammaglobulinaemia has been confirmed and the observation extended to include patients with other types of primary immunodeficiency. Patients with sex-linked congenital hypogammaglobulinaemia have values for mononuclear cell 5'-nucleotidase activity which are in the normal range, whereas most cases of non-familial adult onset `variable' primary hypogammaglobulinaemia have clearly subnormal values. The three patients with isolated IgA deficiency who were tested also had subnormal values. Evidence that the measured enzyme activity is in fact 5'-nucleotidase and independent of interfering phosphatase activities is presented. No significant or consistent alterations in the activities of the following enzymes were detected in mononuclear cells or erythrocytes: adenosine deaminase, purine nucleoside (inosine) phosphorylase, hypoxanthine phosphoribosyltransferase, adenine phosphoribosyltransferase, phosphoribosylpyrophosphate (PRPP) synthetase. The erythrocyte PRPP content and the mononuclear cell PRPP amidotransferase activity were normal in the small number of patients in which they were measured. These findings are discussed in the light of the current interest in the inter-relationship between some disorders of purine metabolism and the immunological deficiency syndromes.     

Chromosome instability in lymphocytes from patients with celiac disease

Cytogenetic studies were performed in celiac disease (CD) patients to determine if the presence of chromosome instability is related to the predisposition to cancer. Chromosome aberrations (CA) and sister chromatid exchange (SCE) frequencies in peripheral blood lymphocyte cultures from untreated CD patients and healthy controls were analyzed. Patients showed aberrations in 23% of cells, while only 3% were detected in the control group (p < 0.0001). The mean frequencies of gaps, breaks and total CA were found to be higher in CD patients compared to controls (p < 0.0001). Breakpoint distribution was nonrandom among chromosomes from celiac patients (p = 0.01), but not among controls (p = 0.04). The frequency of SCE/cell showed a mean value of 6.9 ± 0.6 in CD patients and 7.3 ± 0.2 in controls. No statistical differences were found. Breakpoints involved in CD patients presented a strong coincidence with the location of fragile sites (78.6%) and sites of cancer chromosome rearrangements (57.1%), most of them (75%) associated with malignant non-Hodgkin lymphomas. These results suggest that CD is a condition with increased chromosome instability characterized by a high level of CA and normal SCE frequencies, probably related to the increased incidence of cancer.     

T lymphocytes in synovia of patients with rheumatoid arthritis

Conclusions The observation that rheumatoid synovial-derived lymphocytes display clonal dominance may have important implications in the understanding of the underlying pathogenic processes in RA. Isolation of the relevant T cell clones will allow further characterization including the development of anti-clonotypic mAb which could be used for diagnostic and possibly even for therapeutic purposes. Functional analysis of the clones could be used as an approach to identify the antigen(s) that trigger the disease.Our observations, however, raise a number of points. First, the detection of clonal dominance in a T cell population within a tissue or in blood has been considered a marker of malignancy [9, 13]. Recent studies of several skin diseases suggest that T cell oligoclonality observed in some of these lesions represents a selected repertoire of responding T cells; our study shows that T cell clonality is not restricted to lymphoproliferative diseases but may indeed be a feature of certain inflammatory processes as well.     

Transformation of lymphocytes in patients with certain bacterial infections

The state of cell-mediated immunity was measured by the morphological method of lymphocyte transformation with and without PHA, in a group of 56 patients with bacterial infections and in a group of healthy controls. The patients were divided into three subgroups according to the aetiology: 1. Patients with Gram-positive infections, 2. Patients with Gramn-egative infections, and 3. mixed infections. The transformation values without PHA were significantly (t-test) higher in the Gram-positive subgroups than in the control group. Transformation in cultures stimulated by PHA was much higher in the subgroup of patients with Gram-positive aetiology; the difference between the values in the Gram-negative and in the Gram-positive subgroups was also significant. A decrease of transformation value below 50% was observed in 6 out of 18 patients with Gram-negative aetiology, whereas in the Gram-positive group it was noted in only 2 out of 21 patients.     

Interleukin 2 responsive lymphocytes in patients with adenosine deaminase deficiency

We evaluated the effects of recombinant interleukin 2 (IL-2) on the proliferative responses to mitogens of peripheral blood mononuclear cells (PBMC) from three adenosine deaminase (ADA)-deficient patients. There was significant enhancement by IL-2 of the proliferative responses to phytohemagglutinin (PHA) and pokeweed mitogen (PWM) of PBMC from all three patients. We found that normal PBMC respond with increased numbers of CD3-positive cells when exposed to PHA or PWM and that the response by normal CD8-positive cells was greater than that by CD4-positive cells. In contrast, we found that in ADA-deficient cells the response is almost entirely due to the CD3/CD4-positive population of lymphocytes. These results could not be explained by either the culture conditions or the possibility of a mixed chimeric state. When we evaluated an in vitro cell model of ADA deficiency using an ADA inhibitor, erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA), we found that the inhibitory effect of EHNA plus deoxyadenosine on mitogen-stimulated PBMC could not be prevented by IL-2. These results suggest that the immunodeficiency in ADA deficiency includes the absence or failure of a subset of T cells to make IL-2 and the failure of the CD8-positive subset to respond to IL-2. Also, the in vitro cell model of ADA deficiency using EHNA as the ADA inhibitor is limited in its use in understanding the pathogenesis of this disease.     

Sister chromatid exchanges in lymphocytes of patients with oral carcinoma

Sister chromatid exchanges (SCEs) were studied in cultured peripheral lymphocytes of 22 untreated patients with squamous cell carcinoma of the oral cavity and 29 age- and sex-matched controls. The SCE rate in cancer patients was not significantly higher compared with that found in controls, but there was a significant correlation between the SCE rate in lymphocytes of the cancer patients and the size of the primary tumor.     

T and B lymphocytes in patients with pulmonary tuberculosis.

Counts of T and B lymphocytes were made in the peripheral blood from patients with active and inactive pulmonary tuberculosis and in healthy subjects. In patients with active pulmonary tuberculosis the numbers of T cells were significantly lower than in the controls. After treatment for two months, if the patient's condition improved the percentage of T cells returned to nearly normal levels. If the patient failed to improve, the numbers of T cells remained low. The mechanism of these phenomena is discussed.     

The in vitro proliferation of lymphocytes from patients with hypogammaglobulinaemia

The 5-day cultures from eight patients with primary hypogammaglobulinaemia and from ten normal subjects were compared with regard to the proliferation response to phytohaemagglutinin (PHA), staphylococcal filtrate, streptolysin O and various antigens. No difference was observed between the response of lymphocytes from normal and hypogammaglobulinaemic subjects. A marked proliferation response to at least one antigen was observed in at least five of the eight patients with hypogammaglobulinaemia.     

Radiosensitivity of lymphocytes from patients with Alzheimer's disease

The response after γ-irradiation of lymphocytes from 8 patients with Alzheimer's disease (AD), 3 patients with Huntington's disease and 13 normal subjects to stimulation by phytohaemagglutinin (PHA) was assayed by incorporation of [³H]thymlidine. The response of non-irradiated cells was found to be significantly lower in AD cells than in age-matched normals but not significantly lower in old normals than in young normals. However, the response of irradiated cells to PHA, expressed as a percentage of that in non-irradiated cells, was found to be similar in AD patients, young and old normals and in HD patients.     

Impaired responsiveness of lymphocytes in patients with systemic lupus erythematosus

Cellular immune responsiveness, as measured by lymphocyte transformation in one-way mixed leucocyte cultures (MLC) and in phytohaemagglutinin (PHA) stimulated cultures was evaluated in forty patients with systemic lupus erythematosus (SLE) and in seventy-four normal controls. The effect produced by sera from these subjects on in vitro lymphocyte reactivity was tested on autologous cells and on homologous responding cells from a constant panel of ten healthy volunteers.

The reactivity of lymphocytes from SLE patients to PHA and to a battery of allogeneic cells was significantly lower than that of normal controls.

Sera from some SLE patients inhibited the MLC reactions, while in other cases a distinct stimulatory effect was found.

It is suggested that virus-induced modifications of normal histocompatibility antigens cause the appearance of blocking antibody that might bind to the surface of T lymphocytes, impairing their function.

     

T and B lymphocytes in patients with enteric fever.

Estimation of T and B lymphocytes was done in 50 patients of enteric fever, 50 duration matched non enteric fever patients and 50 normal healthy individuals. The difference in both early and late rosette forming T lymphocytes was found to be statistically significant in enteric versus non-enteric patients. Significant difference was also observed in enteric versus normal individuals in case of late rosette forming T lymphocytes.     

C-band pattern in lymphocytes of patients with soft tissue sarcomas

The pattern of heteromorphisms in the C-band-positive constitutive heterochromatin of human chromosomes No. 1, 9, and 16 was studied in peripheral lymphocytes of 45 patients with soft tissue sarcomas and 78 control individuals. The parameters of the heterochromatic regions analyzed were relative size, symmetry-asymmetry within homologous pairs, and incidence of inversions. No consistent differences were found in these parameters between controls and sarcoma patients.     

Histamine receptor--bearing peripheral T lymphocytes in patients with allergies.

The effect of histamine on the capacity of T lymphocytes to form E rosettes was tested in 10 healthy subjects and in 13 patients with allergies. Histamine had no effect on the capacity of T lymphocytes to form E rosettes in healthy subjects, but significantly inhibited the E rosette formation of T lymphocytes in patients with allergies.