Enhanced endothelial cell proliferation in acute Kawasaki disease (muco-cutaneous lymph node syndrome)

Paired sera from 30 patients with muco-cutaneous lymph node syndrome (Kawasaki disease) were studied for possible effects on human vascular endothelial cells growth in vitro. The majority of sera from acute phase muco-cutaneous lymph node syndrome patients significantly enhanced endothelial cell proliferation more than those from convalescent phase patients, infectious diseases patients, and age-matched normal controls. This stimulation was considered to be specific for EC since muco-cutaneous lymph node syndrome sera did not enhance fibroblast growth more than normal sera. Fractionation of the serum with gel filtration failed to clearly detect the molecular properties of this effect, although both heavy and light material possessed this activity. Extensive search for circulating immune complex in muco-cutaneous lymph node syndrome sera were negative, suggesting that the enhanced endothelial cell proliferation was due to serum components other than immune complexes.     

Possible Role of Streptococcus pyogenes in Mucocutaneous Lymph Node Syndrome IX. Quantitation by ELISA of Streptococcal Pyrogenic Exotoxin in the Serum of MCLS Patients

In the present paper we describe the use of an enzyme-linked immunosorbent assay reinforced with an introduction of monoclonal antibody, for the detection and quantitation of streptococcal pyrogenic exotoxin (SPE) in the serum of patients with mucocutaneous lymph node syndrome (MCLS). The amount of SPE was usually at a high level, and its 100% incidence in patients' sera was proved whenever the assay was made on the day of admission, thereby showing a marked contrast to carefully matched control sera which failed to mediate any positive result. As for the change in detected amount of the toxin, a clear dichotomy was observed between the serum of gammaglobulin-treated patients and that of infants given aspirin; in the former the positive result turned to negative rapidly following the initiation of treatment coupled with a defervescence, while in the latter the reduction of SPE levels was scarcely monitored for as long as 17 days after the onset of illness. Quantitation of SPE might be an auxiliary test for the diagnosis of MCLS, because a considerable amount of SPE was assessed in a patient who developed characteristic huge coronary artery aneurysms following an illness which did not fulfill the diagnostic criteria. These findings support our speculation in relation to the certain role of S. pyogenes as an etiological agent for MCLS. The possible mechanisms of gammaglobulin treatment in reducing the prevalence of cardiovascular lesions and the duration of systemic inflammation are discussed.     

Circulating immune complex in the mucocutaneous lymphnode syndrome

In 16 patients with mucocutaneous lymph node syndrome (MCLS) during the first 2 weeks after the onset (acute phase) and 1 month after the onset (remission phase), measurement of the circulating immune complex (CIC) was performed by a C1q-binding assay (C1q-B.A.) and/or a Protein-A precipitation test (protein-A P.T.). Seven out of 12 samples and four out of nine samples were shown to have raised levels of CIC in the acute phase with the C1q-B.A. and Protein-A P.T. test. In the remission phase, on the other hand, positive results were found in one out of six samples with the C1q-B.A. test and in three out of nine samples with the Protein-A P.T. test. High levels of CIC and disease activity were parallel. Our finding of a higher incidence of positive CIC in MCLS supports the possibility of the immunopathological mechanism.     

Studies on Fc Receptor Bearing T Cells in Infectious and Immunological Diseases of Children: Part 2. Fc Receptor Bearing T Cell in Bronchial Asthama and Renal Diseases

Receptor for the Fc portion of IgG (FcR) has been found on a small population of T cells. The immunological role of these T cells bearing the FcR (Ty cells) remains to be established, but the possibility to be suppressor and/or killer has been suggested. In pathogenesis of atopic disorders, several authors reported the immunodeficiency state. The increase of serum IgE levels may be considered to be the result of low suppressor T cell activity for IgE production. Also, some immunodeficient state may relate to pathogenesis of acute glomerulonephritis (AGN) or nephrosis. In the present study, in order to investigate the immunological pathogenesis of bronchial asthma, AGN and nephrosis, T cells and Ty cells in peripheral blood were counted and the ratio of Ty cells to total T cells was estimated. Then, the significance of these cells in the diseases was discussed. T cells showed slight decrease in asthmatic children examined during attack. The ratio of Ty cells showed no statistical correlation to serum IgE and IgG. In AGN, Ty cells in-creased and showed no change in nephrosis. Although the change of T and Ty cells in bronchial asthma and AGN was revealed, the accurate role of these cells in the pathogenesis is probably complicated and still remains obscure. The change mentioned above may be the reflect of abnormal immune response which is associated with the pathogenesis of these diseases.     

Possible Role of Streptococcus pyogenes in Mucocutaneous Lymph Node Syndrome. XV. Potential Utility of Streptococcal Pyrogenic Exotoxin Toxoid for the Prophylaxis and Treatment of MCLS

Mice made tolerant to streptococcal pyrogenic exotoxin (SPE) by neonatal inoculation with SPE emulsified in incomplete Freund's adjuvant demonstrated early thrombocytopenia followed by thrombocytosis. This state is the perfect counterpart of patients with mucocutaneous lymph node syndrome (MCLS). We have hypothesized that by inducing tolerance to SPE, the biological activities of the toxin might play leading roles in the pathogenesis of MCLS. In the present investigations, the efficacy of SPE on the prophylaxis and treatment of diseases caused by Streptococcus pyogenes (including MCLS) were monitored using the murine model system accompanied with a platelet-counting technique. The mice, rendered tolerant due to neonatal SPE inoculation and followed by immunization with SPE toxoid about 1 month prior to the provocative injections with SPE, demonstrated an almost complete lack of response to the provocation, keeping platelet counts within the normal range of values (except for a marginally significant thrombocytosis 7 days postprovocation). Moreover, anti-SPE titers of the sera from the mice sacrificed on day 35, at which point the observation was terminated, were proved to be markedly elevated when compared with controls. These findings seem to suggest that immunization with the toxoid could overcome tolerance, resulting in the production of an antitoxin. In a second experiment that examined the effect of administration with rabbit antiserum raised against the toxoid, the antiserum-treated mice demonstrated a transitory thrombocytosis on 7 days postprovocation with SPE, followed by an abrupt decrease in the number of platelets from day 10 onward. Such a finding was in complete agreement with that observed in tolerant mice administered with antiserum specific for SPE, suggesting a strong neutralizing activity against SPE of the antitoxoid serum. A third experiment evaluated the effect of F(ab')₂ fragments of the rabbit antiserum to the toxoid on platelet activity. The tolerant mice passively administered i.v. with the F(ab')2 fractions demonstrated a complete lack of responsiveness, except for a small thrombocytosis on days 7 and 10.     

Hemophiliac immunodeficiency: influence of exposure to factor VIII concentrate, LAV/HTLV-III, and herpesviruses

The relationship between hemophiliac immunodeficiency and exposures to factor VIII concentrate, LAV/HTLV-III retrovirus, and infection with Epstein-Barr virus and cytomegalovirus was examined. Exposure to factor VIII concentrate was significantly correlated with decreased percentages of T helper/inducer cells, decreased T helper/suppressor cell ratios, and decreased proliferative responses to plant mitogens. LAV/HTLV-III seropositivity was the primary predictor of increased percentages of HLA-DR-bearing mononuclear cells and decreased proliferative responses to pokeweed mitogen. Epstein-Barr virus and cytomegalovirus infections acted in a synergistic manner with LAV/HTLV-III to produce immunoregulatory defects. Increased percentages of T suppressor cells and decreased delayed cutaneous hypersensitivity skin test responses were observed in LAV/HTLV-III seropositive hemophiliacs infected with Epstein-Barr or cytomegalovirus. We conclude that hemophiliacs receiving commercial factor VIII concentrate experience several stepwise incremental insults to the immune system: alloantigens in factor VIII concentrate, LAV/HTLV-III infections, and herpesvirus infections.     

人类微小病毒B19感染后肾脏疾病

人类微小病毒B19(PVB19)在红系细胞中增殖最佳,并与多种疾病相关。可以检测血清中的特异抗体或用PCR检测其DNA来确定PVB19感染。PVB19感染与肾脏疾病可能相关,此类肾脏疾病有女性趋向性及自限性;多数患者有低补体血症及免疫复合物沉积。该病毒直接破坏血管壁以及免疫复合物的沉积可能为其发病机制。PVB19感染后肾脏病主要是对症治疗,免疫力低下的患者可以静脉注射免疫球蛋白治疗。     

Possible polyclonal B cell activation in mucocutaneous lymph node syndrome

Immunoserological studies on polyclonal B cell activation were carried out on 39 patients with mucocutaneous lymph node syndrome (MCLS) and in age-matched healthy individuals. The incidence of anti-mite, P. acnes (Kato) and EB virus antibodies, recently proposed as aetiological agents by some investigators, was increased in the patient group. Serum immunoglobulin (Ig) M level and IgM-anti-dinitrophenyl (DNP) antibodies, which are considered to be parameters of polyclonal B cell activation, were determined in MCLS cases. The level of serum IgM in MCLS was significantly elevated (0.02<P<0.05). Levels of anti-DNP antibodies in seven cases of MCLS (18%) were significantly higher than those of the controls (P<0.01). Nine of the ten pair sera in MCLS showed a stage-dependent decrease in anti-DNP antibodies. These results suggest that polyclonal B cell activation occurs in MCLS.Abbreviations MCLS mucocutaneous lymph node syndrome - P. acnes Propionibacterium acnes - EB virus Epstein-Barr virus - D. farinae Dermatophagoides farinae - PBS phosphatebuffered saline - Ig immunoglobulin - RT room temperature - FITC fluorescein isothiocyanate - DNP dinitrophenyl - BSA bovine serum albumin - OD optical density     

Tubuloreticular structures and parallel tubular arrays in the lymphocytes of children with aplastic anemia and acute febrile mucocutaneous lymph node syndrome: Electron microscopic observations

Tubuloreticular structure was found in 3 and 30% of the lymphoid cells from two consecutive cases of MCLS and in 10 and 15% of the lymphoid cells from two consecutive cases of aplastic anemia. Parallel tubular array was found in 8 and 24% of the lymphoid cells from the cases of MCLS and in 3% of the lymphoid cells from the cases of aplastic anemia. The possibility of the viral etiology of the both diseases was discussed. eases was discussed.     

Large number of CD19+/CD23+ B cells and small number of CD8+ T cells as early markers for cow's milk allergy (CMA)

Assessment of activation of immune mechanisms is valuable in the early diagnosis of cow's milk allergy (CMA). The purpose of this study was to evaluate peripheral blood lymphocyte subclasses in children suspected of having CMA and healthy infants in order to detect an early marker for food allergy. Altogether 47 breast-fed infants, aged from 0.4 to 10 months were followed-up prospectively from birth because of atopic heredity. Twenty-three of the infants were healthy and 24 infants had a strong suspicion of and later challenge-proven cow's milk allergy. Leucocyte subsets were determined from peripheral blood mononuclear cells by flow cytometry. In response to a clinical cow's milk challenge, seven infants developed urticaria, 11 infants had eczema, three patients had loose stools, diarrhoea or vomiting and three infants had eczema and diarrhoea, loose stools or vomiting. The total percentage of B cells and also the proportion of B cells bearing a low-affinity IgE receptor as a marker for activation were significantly higher, whereas the percentage of CD8+ T cells was significantly lower in infants with challenge-proven CMA than in healthy controIs. These results imply that infants with active CMA have a defect in regulation of B-cell function. Further, they suggest that imbalance of the ratio of suppressor and helper T cells might be an important factor in the etiopathogenesis of CMA. Our results show that large numbers of activated CD19 B cells and low numbers of CD8+ T cells could be considered as early markers for food allergy since they are already detectable in peripheral blood during the earliest symptoms of CMA.     

Thymic dysfunction in histiocytosis-X

In some patients with histiocytosis-X there is a deficiency of suppressor T cells, which is corrected in vitro by incubation with either crude calf thymic extract or thymostimulin. This finding suggests that a thymus deficiency could be involved in this disease. Low levels of serum thymic factor (FTS) are found in patients with histiocytosis-X. Their plasma contain factors capable of inhibiting biological activity of FTS in vitro. Elucidation of the mechanism by which FTS is inhibited would be helpful in understanding the immunological defect in histiocytosis-X. The presence of evidence of thymic dysfunction and the fact that patients respond to thymic hormone therapy suggest that histiocytosis-X could be due to a primary immunodeficiency syndrome. Thymulin, which stimulates the generation of suppressor T cells, could be of benefit in therapy.     

CD4+CD25+调节性T细胞在免疫性血小板减少性紫癜发病机制中的作用

免疫性血小板减少性紫癜是一种以血小板破坏增加和(或)血小板生成异常为特点自身免疫性疾病,以皮肤、黏膜或内脏出血为主要表现.该病发病机制复杂,目前尚未明确.CD4+ CD25+调节性T细胞为抑制细胞,是具有独特免疫调节功能的成熟CD4+T细胞亚群,在小鼠和健康人体中约占外周血CD4+T细胞的5％ ～10％,占人体外周血单个核细胞的1％ ～2％,其通过多种途径对免疫反应发挥抑制效应,能维持内环境的稳定.CD4+ CD25+调节性T细胞功能紊乱或数目异常是导致自身免疫性疾病的重要因素之一,在免疫性血小板减少性紫癜的发生、发展中发挥重要的作用.该文对CD4+ CD25+调节性T细胞的特征和作用及其在免疫性血小板减少性紫癜发病中的作用的研究进展作一综述.     

Possible Role of Streptococcus Pyogenes in Mucocutaneous Lymph Node Syndrome

Migration inhibitory factor (M1F)-induced activity of patients with mucocutaneous lymph node syndrome (MCLS) to antigens associated with Streptococcus pyogenes infection was compared to that of control populations consisting of children with illnesses not related to streptococcal infections (group A), and of patients with streptococcal pharyngitis (group B), with the following results. 1. Though a consiaerable number of patients in the acute state of MCLS failed to respond to antigens consisting of (1) a coccus preparation of S. pyogenes (Picibanil), (2) streptococcal pyrogenic exotoxin (SPE) and (3) an extract from cells of a virally transformed human B-cell line, a complete restoration of their responsiveness was observed in parallel with the decrease of fever. 2. While almost all patients of group A were refractory not only to Picibanil and SPE but also to the extract from the transformed human cells, some children with infections due to intracellular microbes showed responsiveness to the extract from the transformed cells. 3. In group B, some patients showed a marked responsiveness and the other a complete refractoriness to these antigens throughout the observation period. These results raise the possibility that S. pyogenes may play a principal role in the pathogenesis of MCLS.     

Immunological observations before and after successful treatment of chronic mucocutaneous candidiasis with ketoconazole and transfer factor

A girl, 13 months of age, presented with generalised granulomatous skin, hair and mucosal candidiasis. Her lymphocytes failed to respond in vitro to Candida antigen (CA); the intradermal test with CA was also negative. Serum immunoglobulins, complement components, granulocyte functions (phagocytic and fungicidal), T-cell subsets, mitogenic and allogenic lymphocyte stimulation, natural killer cell activity and immune, interferon production were all found to be normal. No circulating immune complexes were detected.Ketoconazole, an antimycotic drug, 5 mg/kg twice daily for 1 month and 2.5 mg/kg twice daily for another month spectacularly cleared all lesions. Afterwards, 4-monthly injections with transfer factor (TF) were given. Intradermal reactivity to CA was observed after the second TF injection. The lymphocyte responsiveness to CA in vitro became strongly positive 3 months after the last TF injection. The level of CA precipitins in serum, which was very high (11 lines) before ketoconazole treatment, decreased to 4 lines. No serum inhibitor of lymphocyte proliferation to CA could be demonstrated in the patient's serum before or after treatment.This specific CA unresponsiveness was not due to an excess of OKT8+(suppressor) cells; macrophage migration inhibiting factor (MIF) production was normal. The nonresponsiveness might be due to antigenic overload or to suppressor cell induction not demonstrable in the present studies. The child has remained free of lesions during 3 years of follow-up without any further treatment.Abbreviations CA Candida antigen - TF transfer factor - MIF migration inhibiting factor - CMCC chronic mucocutaneous candidiasis - FCS fetal calf serum - PHA phytohaemaglutinin - conA concanavalin A - PBMC peripheral blood mononuclear cells - RIA radioimmunoassay - PWM pokeweed mitogen - PMN polymorphonuclear cells     

State of cellular and humoral immunity in children infected with human immunodeficiency virus

Cellular and humoral immunity was examined in 32 children infected with human immunodeficiency virus, living in the Kalmyk ASSR. For comparison purposes, examinations were also made of 6 healthy children and 11 children afflicted with different diseases, living in the same republic. A dramatic lowering of the absolute content of T helper lymphocytes, an appreciable decrease of the T helper/T suppressor ratio associated with the high level of circulating immune complexes were revealed. In children infected with human immunodeficiency virus, grave immunodepression occurred at the early stages of the pathological process. The authors describe the totality of alterations in the immunological parameters, which may give rise to the generalized forms of opportunistic diseases in children infected with human immunodeficiency virus.     

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.     

Kawasaki syndrome--a new disease?

Kawasaki recognized in 1967 the acute febrile mucocutaneous lymph node syndrome (MCLS) as a well defined entity among a variety of hitherto unidentified atypical exanthems. The etiology is uncertain. There are close relations to infantile polyarteritis nodosa (IPN) which is probably the severe variant of Kawasaki's disease. Histologically it is a generalized necrotizing vasculitis, most probably caused by circulating immune complexes. The disease is supposed to be initiated by various infections in patients with certain predispositions. Considerations about etiology and pathogenesis as well as relations to IPN are mainly discussed theoretically. Therefore it is recommended to investigate the Kawasaki syndrome following a devised protocol.     

SOCS低甲基化在儿童过敏性紫癜Th17/Treg细胞失衡中的作用研究

目的 通过研究细胞因子信号转导抑制因子（SOCS）低甲基化与过敏性紫癜（HSP）患儿Th17/Treg细胞失衡的关系,探讨HSP的免疫发病机制。方法 选取2014年5月至2015年1月32例急性期HSP住院患儿为研究对象,另选取行健康体检的28例儿童作为健康对照组。采用ELISA法检测血浆IL-6水平;流式细胞术检测外周血CD4⁺IL-17A⁺T细胞（Th17细胞）比例、CD4⁺CD25⁺调节性T细胞（Treg）比例和CD4⁺T细胞磷酸化STAT3（pSTAT3）蛋白平均荧光强度（MFI）;实时荧光定量PCR（RT-qPCR）技术检测CD4⁺T细胞SOCS1、SOCS3基因mRNA表达;高分辨率熔解曲线（HRM）分析法检测外周血单个核细胞SOCS1基因外显子2、SOCS3基因5'端非翻译区（5'-UTR）可能的STAT3结合位点CpG岛甲基化水平。结果 与健康对照组比较,HSP组血浆IL-6浓度、CD4⁺T细胞pSTAT3的MFI显著增加;HSP组Th17细胞比例显著上调,Treg细胞比例显著下调（P < 0.05）。HSP组患儿急性期外周血单个核细胞SOCS1 mRNA和SOCS3 mRNA水平均显著高于健康对照组（P < 0.05）;HSP组SOCS1 mRNA及SOCS3 mRNA表达均与Th17/Treg比值呈负相关（P < 0.05）。HSP组患儿急性期SOCS1基因外显子2、SOCS3基因5'-UTR区可能的STAT结合位点CpG岛呈低甲基化,而健康对照组呈完全去甲基化状态。结论 SOCS1、SOCS3基因低甲基化所致其相对表达不足可能是HSP患儿Th17/Treg失衡的因素之一。     

Impaired immune regulation in children and adolescents with hemophilia and thalassemia in Israel

The immune function was assessed in 22 children, adolescents and young adults with asymptomatic hemophilia, and 15 with thalassemia, in Israel. Five patients with hemophilia and two with thalassemia were found to be severely abnormal, having cutaneous anergy, very low T-helper cells, elevated T-suppressor cells, inverted T-helper/suppressor ratio, reduced response to mitogens and antigens, and nonfunctional NK cells. Four of the five hemophilia patients exhibited profound lymphopenia also. Decreased T-helper and mildly elevated T-suppressor cells with inverted T4/T8 ratio were observed in the hemophiliacs as a group. In the severe group, the reduction in T-helpers and T4/T8 ratio was more pronounced. The thalassemics as a group were found to have increased numbers of T-suppressor cells with decreased T-helper cells in those with intact spleen only. Both groups studied were found to have elevated IgG levels and low natural killer (NK) activity and normal response to mitogens. Cutaneous anergy was found to be a reliable indication for severe T-cell dysfunction and may serve as an early indication of impending AIDS. These results indicate that patients with hemophilia and with heavily hypertransfused thalassemia may be at increased risk of AIDS as they approach adolescence.     

Graft-versus-host reaction after blood transfusion in a patient with cellular immunodeficiency: The role of histocompatibility testing

Immune complexes (ICs) participate in the pathogenesis of various diseases and can be shown in 18% of all hospitalized patients (excluding those with infectious diseases) by means of a sensitive method such as the Raji-cell radioimmune assay. However, before this test can be applied to quantify disease activity in renal, connective tissue and neoplastic diseases, it must be recognized that febrile infections of the upper respiratory tract also induce ICs in 86% of all patients. The immune complexes containing microbial antigens can be reduced or removed by a single injection of human immunoglobulin. This is a simple method to distinguish between the immune complexes of different specifities. The resulting removal of some immune complexes may be the explanation for the claimed therapeutic effect of gammaglobulin injection in normogammaglobulinemic patients.Supported by Stiftung Volkswagenwerk