Response of human newborn lymphocytes to alloantigen: lack of evidence for suppression induction

Between 1:120 and 1:180 of human newborn T cells proliferate in limiting dilution cultures with allogeneic lymphocytes or with Ia-bearing monocytic stimulator cells. The proliferating responder cells were derived from both the OKT 4+ and OKT 8+ subsets as determined by immunofluorescence and by thymidine uptake. Five to seven days after an exchange blood transfusion there was a slight increase in the percentage of OKT 8+ T lymphocytes in the recipient's blood. Newborn blood also contains a population of non-T cells which proliferate in the absence of allogeneic stimulator cells. In limiting dilution cultures, the frequency of these spontaneously dividing cells was 1:3125 of mononuclear cells. Our results suggest that the newborn T lymphocyte proliferative response to alloantigen is mature by the time of birth and they provide no phenotypic explanation for the previous report of mixed lymphocyte culture-induced suppression by newborn T cells. The predominance of newborn metaphases in 2-way mixed lymphocyte cultures with adult cells (on which the previous report of suppression was based) is not seen if the non-T (stimulator) cells are irradiated. These results suggest that the data previously interpreted as evidence for suppression arose through proliferation of newborn non-T cells.     

Lymphocyte subpopulations in primary immunodeficiency disorders.

Venous blood mononuclear cells from 42 children with primary immunodeficiency disorders and from controls matched for age were studied for lymphocyte subpopulations by E rosetting, surface immunoglobulin, and a panel of anti T cell monoclonal antibodies (OKT series). In 3 cases of severe combined immunodeficiency (SCID) due to adenosine deaminase deficiency, very few circulating T or B cells were found. The other 7 cases of SCID all had normal or, in 3 cases, very high numbers of circulating B cells, but in 6 of these very few cells showed T cell markers. One child had very high numbers of B cells and T cells with an immature pattern of reactivity similar to that found on common thymocytes. In T cell deficient children no consistent pattern was found, but in those with cartilage hair hypoplasia with immunodeficiency there was a low helper (OKT4) to suppressor (OKT8) ratio and high numbers of circulating OKT10 positive cells. In cases of X-linked agammaglobulinaemia circulating B cells were not found but the pattern of T cell markers was normal. In cases of common variable hypogammaglobulinaemia there was a wide scatter of helper (OKT4) to suppressor (OKT8) cell ratios. Five children were studied before and after treatment with the synthetic thymic hormone preparation TP5. There were appreciable alterations in the pattern of staining with anti T cell monoclonal antibodies in 4 of these cases, but in 1 case only was this accompanied by improvements in clinical and immune function.     

Immune status in two brothers with Omenn's syndrome: no discernible chimerism on FACS analysis using a monoclonal antibody specific for a maternally restricted HLA antigen

Sequential immunologic examinations, including lymph node biopsies, in two brothers with clinical characteristics of Omenn's syndrome are presented in this study. Although the number of circulating T cells with mature phenotype (OKT3+, TCR1+) was within normal range, the lymphocyte proliferative response to mitogens was poor. Examinations of the lymph nodes revealed marked lymphoid depletion associated with eosinophilic infiltration and reticular cell proliferation. Over the clinical course of 5 months, circulating T cells also mostly disappeared. Thymic hypoplasia was noted at autopsy. Although intrauterine graft-versus host disease (GVHD) has been hypothesized as being the pathogenetic mechanism in this syndrome, maternal lymphocytes circulating in these patients were not identified either by karyotype and HLA typing or by highly sensitive FACS analysis and immunohistochemical studies using a monoclonal antibody, HLA-A9, specific for a maternally restricted HLA antigen, Aw24. In conclusion, the familial occurrence and the absence of maternal chimerism might be the essential features of Omenn's syndrome which should be differentiated from fetal GVHD.     

Unusual Lack of the Epitope Recognized by OKT4 on the Helper/Inducer T Lymphocytes

Lack of the epitope recognized by OKT4 monoclonal antibody on the helper/inducer T lymphocytes in a 14-year-old boy with IgA nephritis is described. The lymphocytes reacted normally with OKT3 /Leu4 and OKT8/αLeu2a monoclonal antibodies but not with OKT4 monoclonal antibody. Studies with other monoclonal antibodies (αLeu3a, OKT4A, OKT4B, OKT4C, OKT4D) which also identify the helper/inducer T lymphocyte subset revealed that cells of this population were present in normal numbers among the lymphocytes of the peripheral blood. Staining with OKT4 plus αLeu3a in normal persons indicated that T4 antigen is present on a small population of lymphocytes which lack Leu3a antigen. Further, the intensity of staining of the majority of cells in the subpopulation is increased when these two fluorescienated antibodies are used together. In this patient neither this small OKT4⁺ Leu 3a^- population nor the cells bearing the Leu3a antigen showed OKT4 staining. The findings in the surface marker analysis of E+ OKT8- peripheral lymphocytes which were achieved by panning of the patient's peripheral cells indicated the existence of a population of E⁺OKT8^- peripheral lymphocytes which were achieved by panning of the patient's periphera cells indicated the existence of a population of E⁺ (4A⁺4B⁺4C4D⁺)αLeu3al⁺ ly mphocytes in this patient. Lymphocyte responses to PHA, ConA and PWM, however, were all within normal range. Further, this patient had normal serum immunoglobulin levels and exhibited no symptoms or signs of immunodeficiency. These findings indicate that the patient under study has functionally normal helper/inducer T lymphocytes which lack the epitope recognized by OKT4 monoclonal antibody.     

Long-term study on T lymphocyte subsets in newly diagnosed type 1 diabetes mellitus

T lymphocyte subsets in peripheral blood from 16 newly diagnosed type 1 diabetic children were studied prospectively at four time intervals: as soon as possible after diagnosis and 1, 4 and 12 months later. T lymphocyte subsets were analysed using monoclonal antibodies and counted by cytofluorimetry. The percentage of T lymphocytes (OKT3+ cells) did not change at the four study times. The percentage of helper/inducer T cells (OKT4+ cells) was high at the diagnosis (43.1 +/- 2.1%), but decreased after 1 and 4 months with no difference in the control values. The percentage of suppressor/cytotoxic T lymphocytes (OKT8+ cells) was low at the diagnosis, but increased after 1 and 4 months. The OKT4/OKT8 ratio was 2.31 +/- 0.22 at the diagnosis study, decreasing to 1.83 after 1 month, compared with 16 sex- and age-matched control children. The high percentage of helper/inducer T lymphocytes and low number of suppressor/cytotoxic T cells at onset of diabetes favour immune reactions that lead to beta-cell damage.     

Transient remission after intercurrent measles infection in a patient with hyperimmunoglobulin E syndrome

A 5-yr-old patient with hyper IgE syndrome contracted measles. This was accompanied by a temporary disappearance of his skin lesions. The patient had a long history of recurrent infections, chronic eczematoid pruritic dermatitis, and elevation of serum IgE level since infancy. Immunologic studies revealed decreased suppressor T cells (OKT8 cells) with increased IKT4/OKT8 ratio, defect in suppressor T cell function, and decreased chemotactic index. In February 1985, when he developed an interrcurrent measles infection at age of 5, the eczematoid pruritic dermatitis disappeared completely and immunologic defects improved transiently, with normalization of OKT4/OKT8 ratio, decrease in in vitro IgE synthesis, in vivo serum IgE level, and interleukin-2 production, decreases in IgG Fc receptor-bearing cells and autologous mixed lymphocyte reaction, and normalization of chemotactic index. One month later, the eczematoid skin lesion relapsed and immunologic defects reappeared. These studies suggested that the pathogenesis of hyper IgE syndrome involved a hypofunction of suppressor T cell. The transient remission associated with measles infection is probably related to the effect of the virus on the helper T cells, resulting in a normalization of OKT4/OKT8 ratio and IgE production.     

小儿特发性血小板减少性紫癜血小板相关抗体、T细胞亚群检测及其临床意义

本文对1989年9月至1995年2月收治的112例小儿特发性血小板减少性紫癜的血小板相关性抗体、T细胞亚群检测及其临床意义进行了探讨。结果表明：急、慢型无论是OKT3及OKT4+均低于正常对照组，且有显著性差异，P值均＜0．05；而OKT8+与正常对照组相比则较高，P值＜0．025。但是，急、慢两型相比均未见统计学上显差性差异。本文的检测血小板抗体lgG、IgM及IgA，显示均有所增高尤以PAIgG最明显，阳性率达85．7％，与文献报道近似。经治疗10例血小板动态变化，PAIg随着血小板的恢复均下降至正常，呈负相关.     

Local T cell subsets in mumps meningitis.

In the acute phase of mumps meningitis, more than 85% of the cells in cerebrospinal fluid (CSF) were OKT 3 positive, while 76% of the peripheral mononuclear cells (PMN) were OKT 3 positive. The ratio of OKT 4:8 positive cells in CSF was significantly lower than that in PMN, showing that suppressor/cytotoxic T cells had selectively accumulated in CSF. In addition, 58% of CSF cells were immune associated (Ia) positive, probably activated T cells.     

The human cytolytic T lymphocyte response to transplantation antigens

Cytolytic T lymphocytes are important effectors in the immune response to allografts, viral infections, and some tumors. Cytolytic T lymphocyte lines and clones have been generated and used to define functionally relevant effector and target cell surface molecules. The major target antigens of the human allogeneic response are the major histocompatibility complex antigens. Functionally relevant effector antigens include LFA-1, LFA-2, LFA-3, OKT4, OKT8, OKT3, and Ti, the T cell receptor. The diagnostic and therapeutic implications of T cell surface molecules are discussed.     

The measurement of leukocyte subsets in the peripheral blood of cancer patients with solid tumors using monoclonal antibody reagents

Monocyte and lymphocyte subsets were quantitated in the peripheral blood of normal subjects and patients with solid tumors using the monoclonal antibody reagents OKM1, BRL63D3, OKT3, OKT4, and OKT8. Percentages and numbers of cells reacting with monoclonal reagents were analyzed by indirect immunofluorescence. Correlations between leukocyte subset values and stage of disease or immunocompetence were sought. No differences from normal were seen in the percentage of OKT3, OKT4, and OKT8 cells or in the ratios of OKT4/OKT8 cells in peripheral blood mononuclear cells (PBMC) from all cancer patients, patients with localized disease, or patients with advanced disease. A significant decrease in absolute numbers of lymphocytes, OKT3 cells, OKT4 cells, and OKT8 cells was seen in the peripheral blood of patients with advanced disease reflecting the absolute lymphopenia of these patients. A significant increase was seen in the percentages of PBMC reacting with OKM1 and BRL63D3 from patients with advanced disease compared with normal donors or localized disease patients. A positive correlation was found between PHA responsiveness and absolute numbers of OKT3 and OKT4 cells. A negative correlation was found between PHA responsiveness and percentages of OKM1 cells. These data indicate that malignant disease does not alter T cell subset percentages in patient peripheral blood but may decrease their absolute numbers in association with absolute lymphopenia. On the other hand, percentages of OKM1 and BRL63D3 cells are increased in patients with advanced solid tumors in association with impaired PHA responsiveness.     

Enhanced suppressor T cell activity resulting in increased IgM and decreased IgG productions in children with minimal change nephrotic syndrome

In order to clarify the cellular basis accounting for the decreased serum IgG and increased serum IgM in children with minimal change nephrotic syndrome (MCNS), peripheral blood mononulcear cells (MNC) were obtained from 15 children with biopsy-proved MCNS and 15 age- and sex-matched normals. MNC were then separated into helper (OKT4) T cells, suppressor (OKT8) T cells, B cells and monocytes. Different cell populations from patients and normals were then recombined and pokeweed mitogen-stimulated in vitro immunoglobulin biosynthesis was studied. The results strongly suggest the presence of isotype-specific suppressor T cells which may affect the switch of IgM B cells to IgG B cells in patients with MCNS and may be used to explain partly the clinical findings of lowered serum IgG and increased IgM in those individuals.     

In vitro effect of a bovine thymic extract (thymostimulin) on T-cell differentiation in cord blood lymphocytes

Cord blood lymphocytes were isolated from 40 normal newborns. The initial 20 samples were used to determine the dose-response curve of bovine thymic extract (Thymostimulin) by the measurement of active T cells. Results showed that the active T cells increased significantly when the Thymostimulin concentration was increased to 50 ng/ml and 100 ng/ml. The remaining 20 samples were divided into three portions and preincubated either with 50 ng and 100 ng of Thymostimulin or without Thymostimulin. The total T cells, active T cells, B cells, T-cell subsets, and lymphoproliferative responses to phytohemagglutinin, concanavalin A, and pokeweed mitogen were determined. The results showed that the active T cells, total T cells, B cells, OKT4 cells, and OKT8 cells were significantly increased after Thymostimulin treatment. The lymphoproliferative responses were also significantly increased. These data strongly support our conclusion that Thymostimulin has a marked stimulating effect on human lymphocyte proliferation and differentiation.     

T cell subsets in human colostrum

Lymphocytes from 10 paired colostrum and peripheral blood specimens were examined to determine if the colostral T cell population differs from the peripheral blood T cell population in subset distribution. The percentages of lymphocytes staining with OKT3, OKT4, and OKT8 murine monoclonal antibody were determined. Lymphocytes from colostrum were 74.7 +/- 2.5% OKT3+, 50.6 +/- 2.3% OKT4+, 24.0 +/- 1.7% OKT8+, whereas peripheral blood lymphocytes were 78.7 +/- 1.9% OKT3+, 48.4 +/- 1.4% OKT4+, and 29.8 +/- 1.6% OKT8+. The percentage of colostrum lymphocytes positive for OKT3 was significantly although not strikingly lower than the OKT3 percentage for blood lymphocytes (p less than 0.05). This difference was due to the lower percentage of OKT8 positive lymphocytes in colostrum compared with blood (p less than 0.01). Although the T cell subset distribution of colostrum generally appears to be similar to that in the peripheral blood, there were small differences in OKT3 and OKT8 percentages that were statistically significant suggesting the possibility of some selectivity of the colostral T cell population.     

T cell subsets in glomerulonephritis

Abnormal lymphocyte function has been postulated to have a pathogenetic role in nephrotic syndrome. In an attempt to investigate the pathogenetic role of lymphocyte subsets in human glomerular disease, we studied 110 children suffering from nephritis during the acute nephrotic phase or nephritis without steroid treatment, 4 weeks later after steroid treatment, in remission and relapse. These patients included minimal change nephrotic syndrome (MCNS) 15 cases, focal segmental glomerular sclerosis (FGS) 6 cases, mesangial cell proliferative nephropathy (MesPGN) 42 cases, membranoproliferative glomerulonephritis (MPGN) 2 cases, hepatitis B surface antigenemia associated with membranous nephropathy (HBVMN) 10 cases, IgA mesangial nephropathy (IgAN) without nephrotic syndrome 7 cases, poststreptococcal glomerulonephritis (PSGN) 24 cases and chronic glomerulonephritis (CGN) 4 cases. There was no significant difference in the total lymphocyte count of each different pathological group of nephritis except that lymphopenia was noted in the CGN patients. When the lymphocyte phenotypic profile was examined, OKT8 cells were significantly increased in the MesPGN patients and both OKT4 and OKT8 cells were significantly increased in HBVMN. Comparison of MCNS and MesPGN during the acute nephrotic phase showed the OKT4/OKT8 ratio decreased significantly in MesPGN. Four weeks after steroid treatment, OKT4 cells decreased both in MCNS and MesPGN being pronounced in MCNS. In the remission stage with steroid treatment the OKT4/OKT8 ratio decreased in MCNS and was mildly elevated in MesPGN. In relapse, the OKT4/OKT8 ratio was the same as it was during the onset of nephrotic phase. MCNS cases were steroid responsive whereas in MesPGN there were frequent relapses or partial steroid response.(ABSTRACT TRUNCATED AT 250 WORDS)     

Defective regulation of the protective IgE response against intestinal helminth Ascaris lumbricoides in malnourished children

It is well established that malnutrition affects the immune response and increases the susceptibility to parasitic infection. In the present study we evaluated some aspects of the cellular and cytokine network that regulate the IgE responses, which are important components of host defence mechanisms against helminthic parasites in children infected with the intestinal helminth Ascaris lumbricoides, and with differing degrees of malnutrition. We found a defective T cell response in malnourished children, as indicated by diminished levels of circulating total (CD3+), helper (CD4+), IL-2-receptor-bearing (CD4+CD25+) and memory helper T cell responses (CD4+CD45RO+) in keeping with the decreased specific IgE levels against Ascaris lumbricoides. In contrast, the proportions of total B cells (CD20+), and those bearing the low-affinity IgE receptor (CD23+) were increased in the moderated malnourished children. Moreover, serum IL-4 levels and total IgE were also increased in these children. We suggest that malnutrition can cause an imbalance in T cell subpopulations that may lead to a defective T cell maturation and a decreased specific anti-Ascaris IgE response thus increasing the susceptibility to such infections. The high levels of total IgE observed may be related to a non-specific stimulation of the proliferation of activated B cells, probably caused by helminthic parasites and other infectious agents that are frequent in malnourished children.     

Acute Infectious Lymphocytosis: Phenotype of the Proliferating Cell

ABSTRACT. A case of acute infectious lymphocytosis in an otherwise healthy 2-year-old child is reported. Marker analysis of the expanded blood lymphocytes showed that they were predominantly T cells and that there was a considerable increase in the helper/inducer phenotype (OKT4⁺) population. However, the lymphocyte response to polyclonal T-cell activators was low. This is the first report on T-cell subset distribution in acute infectious lymphocytosis.     

Childhood common variable immunodeficiency with autoimmune disease

Clinical and laboratory findings in eight patients with childhood common variable immunodeficiency and autoimmune disease are described. Six of the eight patients had initial signs of the disease, persistent secretory diarrhea, recurrent upper respiratory tract infections, or both, in the first year of life. Autoimmune manifestations included idiopathic thrombocytopenia (4/8), hemolytic anemia (3/8), secretory diarrhea (4/8), arthritis (2/8), chronic active hepatitis (2/8), parotitis (2/8), and Guillain-Barré syndrome (2/8). In addition to the expected sinusitis, otitis, and pneumonia caused by encapsulated bacteria, these patients also had severe infections with viruses of the herpes group. Most of these patients had lymphadenopathy, splenomegaly, growth failure, and failure to develop secondary sexual characteristics. Laboratory studies demonstrated a significant increase in the ratio of T cells expressing the T helper phenotype (OKT4) to T cells expressing the T suppressor-cytotoxic phenotype (OKT8) (T4/T8). This increase could be attributed to a decrease in the absolute number of T8 cells. Additional findings included fluctuating levels of serum immunoglobulins and markedly diminished in vitro antibody production by B cells. The clinical course was relapsing and remitting, and dominated by the autoimmune manifestations of the disease. This group of patients constitutes a distinct subset of children with hypogammaglobulinemia, a subset with a complex, multisystemic disorder associated with significant morbidity and mortality.     

OKT3 treatment of steroid-resistant rejection in pediatric liver transplant recipients.

Of a total of 187 consecutive liver grafts in 149 pediatric recipients, 59 episodes of steroid-resistant, biopsy-proven rejection (32% of grafts) were treated with OKT3 monoclonal antibody. After 59 OKT3-treated episodes, liver function at the end of treatment was normal in 40%, improved in 35%, and unchanged in 24%. Of 21 partial responses, 12 episodes eventually resolved to yield an overall complete response rate of 59%. CD3-positive T-cells greater than 5% occurred during 61% of OKT3-treated rejection episodes and was associated with impaired efficacy of OKT3 (30% complete response rate). Six grafts were re-treated with OKT3 for rejection and in all CD3-positive T-cells could not be maintained less than 5%. Of the six grafts requiring repeat OKT3, five failed-retreatment and required retransplantation. OKT3 antibodies of low titer were found prior to a second use of OKT3 in 65% of episodes. Patients treated with OKT3 after failing more than two preceding steroid courses had a significantly increased chance of graft loss (57%; p = 0.01). We conclude that this group of pediatric patients appeared less responsive to OKT3 compared to other series combining pediatric and adult recipients, possibly due to a more vigorous immune response in the child.     

The immunological profile of children with Down's syndrome

In 1965 Benda demonstrated that bioptic and autoptic material of children with Down's syndrome showed hypoplasia of the thymus with poor histological differentiation between the cortex and medulla and impairment of Hassal's corpuscles which were also fewer than normal. In the seventies studies revealed the increased susceptibility to infections and higher incidence of leucosis in Down's syndrome patients as well as changes of immunologic defences (in particular cell mediated immunity). This study examines 35 children (16 boys and 19 girls) aged 6 months-20 years. Subjects were divided into a group of 18 cases in poor health with a history of recurrent infections and a group of 17 children in good health. Skin tests were performed by inoculating 0.1 ml of a solution formed by 1 ml physiological solution and 0.1 ml tetanus toxoid. Skin reaction was evaluated 48 hours later. Lymphocyte typing tests were performed with the rosette method and with monoclonal antibodies for T lymphocytes and with the determination of surface immunoglobulins for B lymphocytes. OKT4 (T helper), OKT8 (T suppressor) subsets assayed and the OKT4/OKT8 ratio was determined. Skin tests were negative in 3 cases (8.6%). The number of B lymphocytes was normal in all children. Total number of lymphocytes was decreased in 51.4% of cases. Two subjects had a reduction of OKT4 and 14 had an increased of OKT8 and 16 a significantly lower OKT4/OKT8 ratio. It is clear that skin tests were normal also in those children with low total lymphocyte values. The most closely related parameter to mobility was the OKT4/OKT8 ratio and the most distantly related on was the skin test. Only 3 cases had modifications of all 3 parameters together. Apart from the constant and complete immunological deficit described by many authors and which we cannot confirm the results of this study are in agreement with those of other authors.     

Sequential fine needle aspiration biopsy in glomerulonephritis

Cellular immune mechanisms have been shown to play a prominent role in glomerulonephritis. Cellular mediators of inflammation cause both acute and progressive glomerular and tubular injury. Understanding the mediation pathways offers the opportunity for therapeutic manipulation. In addition to polymorphonuclear leucocytes, monocytes/macrophages, B-cells and T-cells subsets are being enumerated in normal and diseased renal tissues. The correlation between immunological findings in peripheral blood and infiltrate composition in renal tissue, by using weekly Fine Needle Aspiration Biopsy (FNAB), for assessing the clinical status and monitoring the immunosuppressive therapy was the aim of this study. When determining the intensity of inflammation the numerical values of the Total Corrected Increment (T.C.I.) were defined as follows: less than 1.5 no inflammation; from 1.5 to 2.0 inflammation possible; greater than 2.0 inflammation. The ratio between OKT4 and OKT8 was used as the index: greater than 2.0 immunologic activation; greater than 2.0 no immunologic activation. When the T.C.I. was greater than 1.5 and the OKT4/OKT8 less than 2.0, or the T.C.I. less than 1.5 and the OKT4/OKT8 greater than 2.0 we used only a standard treatment. When both activation indexes were in the normal range we have not treated the patients. When the T.C.I. was greater than 1.5 and the OKT4/OKT8 was greater than 2.0 we treated the patients with standard treatment plus methylprednisolone pulses every time the activation indexes monitored by FNAB, showed an increase. A spontaneous improvement was obtained in untreated patients. The patients treated by standard therapy alone showed a different outcome. All patients treated with standard therapy plus methyl-prednisolone pulses showed a progressive clinical improvement.(ABSTRACT TRUNCATED AT 250 WORDS)