In vitro B-lymphocyte switch disturbance from IgM into IgG in IgM mesangial nephropathy

In vitro immunoglobulin (Ig) synthesis using a co-culture technique after activation of lymphocytes with pokeweed mitogen, T-cell subsets and interleukin-2 (IL-2) production was studied in 10 children who suffered from IgM mesangial nephropathy (IgMN), 10 children who suffered from minimal change nephrotic syndrome (MCNS) with hypercellularity and 6 children who suffered from MCNS with normal cellularity during the acute nephrotic phase. Reduced in vitro IgG production was found in the presence of OKT8 cells from all groups of patients. However, in vitro IgM production was increased only in OKT8 cells from IgMN and MCNS patients with hypercellularity. In vitro Tac expression on the OKT8 cells, IL-2 production, T-cell subsets including Leu2a⁺15⁺ (suppressor T-cells), Leu2a⁺DR⁺ (activated suppressor T-cells) and Leu3a⁺8⁺ (suppressor T-cell inducer) were all increased in IgMN and MCNS patients with hypercellularity. There was a significant correlation between in vitro IgM production by co-culture technique and IL-2 production. These results strongly suggest the hyperfunction of isotype-specific suppressor T-cells which may affect the switch of IgM B-cells to IgG B-cells in IgMN and MCNS patients with hypercellularity and may be used to explain in part the clinical findings of lower serum IgG and increased IgM in those patients.     

小儿特发性IgM肾病与微小病变肾病综合征的临床病理研究

对于ＩｇＭ在肾小球系膜区沉积的临床和病理意义争论颇多，我们就２５例ＩｇＭ肾病（ＩｇＭＮ）与１６例微小病变肾病（ＭＣＮＳ）患儿的临床与病理进行对比分析，结果表明ＩｇＭＮ与ＭＣＮＳ临床表现极为相似。但ＩｇＭＮ对肾上腺皮激素治疗反应不佳，且易反复或复发。这可能与ＩｇＭＮ的病理改变较重有关。另外作者还对ＩｇＭ在系膜区沉积潜在临床与病理意义进行了初步探讨，认为ＩｇＭ在系膜区沉积无明确的临床和病理意义。     

Histopathological and immunological studies in spontaneous remission of nephrotic syndrome after intercurrent measles infection

Two cases of childhood nephrotic syndrom (NS) were diagnosed in September 1981 and February 1982, respectively. The patients were first cousins. There was no other family history of renal disease. Five months after onset of nephrosis in 1 patient and 1 month after onset of nephrosis in the other both had an intercurrent measles infection and the NS rapidly went into spontaneous remission. However, relapses occurred 3 and 7 months later. At that time both patients were started on steriod treatment and obtained remission. Unfortunately, both relapsed again 14 months later. Kidney biopsies were performed, the pathologic findings demonstrated that both were mild mesangial proliferative glomerulonephritis. The mononuclear cell subsets and lymphoproliferative responses were studied during the acute measles infection, 4 weeks later, in remission and relapse of NS. The mononuclear cell subsets and lymphoproliferative response in medium containing autologous serum with complement were decreased during the acute measles infection. Both patients had an increase of OKT8 cells and Leu-7 cells in relapse and a decrease in remission. Taken together, natural measles infection caused a prolonged depression of cell-mediated immunity, T cell subset and induced a temporary remission of steroid-sensitive NS.     

14例儿童IgM肾病临床与病理探讨

目的：探讨儿童IgM肾病临床与病理之间的关系。方法：对首都儿科研究所附属儿童医院2004年10月～2009年3月经肾组织活检明确诊断为IgM肾病14例患儿的临床资料、肾组织病理、治疗方法和随访情况进行回顾性分析。结果：本资料50%IgM肾病临床表现为肾病综合征;肾脏病理显示以系膜增生病变为主,免疫荧光示8例单纯IgM沉积,5例IgM＋IgG沉积;结合临床和病理给予个体化治疗,有效率可达81.8%。结论：IgM肾病临床和病理存在着一定的关联性,临床表现为肾病综合征者病理损害较重。早期、规律、个体化治疗,近期疗效好。大量蛋白尿是预后不良的因素。     

Serum IgE in primary glomerular diseases and its clinical significance

Total serum IgE was measured in 119 cases of primary glomerular diseases and 33 normal healthy persons. Statistically significant higher levels were noted in minimal change disease (MCD; median: 630 U/ml), IgM nephropathy (IgMN; 618 U/ml), focal glomerulosclerosis (FGS; 373 U/ml) and membranous glomerulonephritis (MGN; 144 U/ml). A higher level of serum IgE was noted in association with more frequent relapse or steroid resistance in MCD and IgMN and in FGS with nephrotic syndrome. A small group of IgA nephropathy with nephrotic range proteinuria was also noted to have extraordinarily high serum IgE. These findings suggest that IgE may play an important role in the pathogenesis of MCD, IgMN, and FGS and may serve as a prognostic indicator in terms of steroid responsiveness in MCD and IgMN.     

T-lymphocyte subsets in nephrotic syndrome.

T-lymphocyte subsets when measured in steroid responsive nephrotic syndrome (SRNS) have demonstrated significant variance from normal values. T-cell subsets were studied by using two-color flow cytometric analysis in 32 children (9.2 +/- 5 years of age) with SRNS. The children were divided into four groups: a) SRNS in acute relapse, on prednisone; b) SRNS in acute relapse, off prednisone; c) SRNS in long-term remission, off prednisone (nephrotic controls); d) patients in remission on long-term prednisone therapy; and e) 15 age-matched normal controls. Children suffering an acute relapse of SRNS showed an increase in Leu2a+/DR+ (CD8) activated lymphocytes (P less than 0.05), a decrease in Leu4a+ total T-lymphocytes (P = 0.01) and a decrease in Leu3a+ (CD4) helper T-cells (P less than 0.05) when compared to normal controls and nephrotic controls. Though some subset changes may represent a prednisone effect and the functional role of these lymphocytes in the disease process is unknown, this study provides additional evidence to support a role for abnormal T-cell subsets in the etiology of SRNS.     

IgM nephropathy: clinical picture and pathological findings in 36 patients

Immunoglobulin M nephropathy (IgMN) is an idiopathic glomerulonephritis characterized by a variable degree of mesangial hyper- cellularity and diffuse IgM deposits. This study describes the clinical presentation and the morphological findings in 36 patients, five adults and 31 children, with IgMN. The initial manifestations of the disease were the nephrotic syndrome in 32 patients, proteinuria in two, hematuria associated with proteinuria in 16 and isolated recurrent gross hematuria in two patients. The nephrotic syndrome was steroid responsive in 9% of the cases, steroid dependent in 25% and steroid resistant in 66%. Five patients were hypertensive at the onset of the disease. The mean follow- up period was 3.4 years (range, 1-7 years). One patient developed end- stage kidney disease five years after the diagnosis. On light microscopy, 24 specimens showed mild focal and segmental mesangial hyper- cellularity, 10 cases were normal in cellularity and two cases showed diffuse global mesangial hyper- cellularity. Four cases showed focal and segmental glomerulosclerosis. Mild interstitial inflammation and fibrosis were observed in 11 cases and focal tubular atrophy and hypertensive hyaline arteriosclerosis were noted in three cases. In addition to IgM, the immunofluorescence study showed C3 deposits in 12 cases, IgG in nine cases and IgA in one case. Electron dense- deposits in the mesangium and para- mesangial areas were demonstrated on electron microscopy in 22 cases. In view of its different clinical presentations and the constant findings of diffuse mesangial IgM deposition in all the cases, it is concluded that this form of nephropathy constitutes a distinct entity separate from minimal change disease or focal and segmental glomerulosclerosis. Patients with IgMN had a higher prevalence of steroid resistance and dependence in the proteinuric group.     

Changes in serum lipoprotein(a) levels in children with corticosensitive nephrotic syndrome

Variations in lipoprotein(a) [Lp(a)] levels were evaluated during the course of the nephrotic syndrome in 20 children (17 males, 3 females, aged 2 – 16 years), to evaluate the use of this parameter in the prognosis and monitoring of the disease. One patient was in relapse, 12 in remission, and 7 alternated between remission and relapse. Results were compared with those obtained in a control population of 100 age-matched children. Lp(a) was measured by a previously described immunonephelometric technique. Serum Lp(a) levels were increased in children with relapsing nephrotic syndrome compared with controls (median value of 419 mg/l vs. 86 mg/l). The median Lp(a) level in patients with nephrotic syndrome in remission was different from controls (270 mg/l under steroid therapy and 163 mg/l without steroid therapy), but remained within the reference range. Of the patients in relapse, 66% had Lp(a) levels above the generally accepted limit for cardiovascular risk of 300 mg/l, compared with 16% of controls, 44% of patients with nephrotic syndrome in remission under steroid therapy, and 18% of patients with nephrotic syndrome in remission without steroid therapy. In 2 patients, Lp(a) was measured repeatedly and was significantly higher during the acute phase of the disease (up to sixfold basal level). Changes in Lp(a) levels correlated with cholesterol levels, but the kinetics and the extent of variations were different. These data suggest that measurement of Lp(a) is useful for monitoring the nephrotic syndrome in children, particularly for detecting complications. Received January 3, 1995; received in revised form and accepted January 31, 1996     

Clinico-pathological characterization of mesangial proliferative glomerulonephritis with predominant deposition of IgM

Background. IgM nephropathy (IgMN) was first described in 1978 but whether IgMN is a distinct entity has not yet been confirmed. We tried to clarify the characteristics of IgMN on the basis of clinical and histological findings of patients who were treated and investigated by us. Methods. We evaluated 90 patients (group A) with mesangial proliferative glomerulonephritis (MesPGN) who exhibited more dominant mesangial deposits of IgM than of IgG and IgA. To clarify the significance of the IgM mesangial deposits, we divided these patients into two subgroups: group A1 comprised 30 patients with diffuse and global mesangial IgM deposits and group A2 comprised the other 60 patients, with focal segmental mesangial IgM deposits. Ninety Mes-PGN patients without predominant IgM deposits, excluding those with IgA nephropathy were studied as controls (group B). Results. Group A had significantly higher levels of serum IgM than group B (P < 0.01). The mean values for glomerular filtration rate (GFR), serum albumin, and CH50 in group A were significantly lower than those in group B (P < 0.01). Proteinuria in group A was significantly more severe than in group B (P < 0.05). Group A had a significantly higher incidence of adhesion (P < 0.01) and duplication (P < 0.05) of capillary loops than group B. They also demonstrated a significantly higher incidence of glomerular deposition of C1q and/or C4,C3, and fibrinogen than group B (P < 0.01). Conclusion. These findings suggest that the mesangial deposition of IgM is useful for assessing glomerular damage or renal dysfunction in MesPGN patients, regardless of the amount of mesangial IgM deposits. Furthermore, MesPGN with predominant IgM deposition, which includes both diffuse global and focal segmental deposition of IgM, should be referred to as IgM nephropathy. Received: October 20, 1997 / Accepted: October 8, 1998     

儿童肾病综合征T细胞亚群检测的临床意义

目的：探讨儿童原发性肾病综合征（NS）T细胞亚群检测的临床意义。方法：对25例NS活动期和缓解期患儿应用流式细胞仪检测T2细胞亚群的变化。结果：NS活动期组CD3^ ,CD4^ ,CD4^ /CD8^ 比值，NK[CD(16 56)]^ 细胞均明显低于缓解期组（P＜0．01）和对照组（P＜0．01）。结论：说明NS细胞免疫功能减低，T细胞亚群检测可作为肾病综合征活动指标之一。     

T-cell subsets,interleukin-2 receptor expression and production of interleukin-2 in minimal change nephrotic syndrome

This study was designed to investigate T-lymphocyte subsets interleukin-2 receptor (IL-2R) expression and IL-2 production in minimal change nephrotic syndrome (MCNS). Peripheral blood T-lymphocytes and IL-2R expression were analysed using fluorescein isothiocyanatelabelled CD3, CD4, CD8 and CD25 monoclonal antibodies with flow cytometry. IL-2 production was determined by enzyme immunoassay. Ten children with MCNS in relapse and in remission were evaluated. Thirteen healthy children served as controls. The patients in relapse demonstrated a moderate decrease in the total absolute lymphocyte counts and CD8(+) T-lymphocytes compared with controls (P<0.05) and had a greatly increased IL-2R expression in frashly isolated, unstimulated peripheral lymphocytes compared with patients in remission and controls. While this was not statistically significant, IL-2R expression on cultured lymphocytes stimulated with phytohaemagglutinin was significantly elevated in relapse compared with those in remission and controls (P<0.05). IL-2 production did not correlate well with IL-2R expression and there was no significant difference between the groups. Our results suggest that T-cell subset changes and high IL-2R expression on peripheral lymphocytes may indicate the presence of stimulated T-cell populations in MCNS which could contribute to the immunopathogenesis.     

Minimal change disease with IgM+ immunofluorescence: a subtype of nephrotic syndrome

Immunoglobulin (Ig) M nephropathy is defined by electron-dense mesangial deposits and mesangial IgM visible by immunofluorescence (IF) without other histopathologic and immunofluorescent microscopic abnormalities. Certain patients have only immuno-positive (IgM+) IF. Children presenting with steroid-dependent or steroid-resistant nephrotic syndrome have a high prevalence of IgM+ IF with or without electron-dense deposits. We reviewed the clinical course of children with steroid-dependent or steroid-resistant nephrotic syndrome who underwent renal biopsy at Texas Children‘s Hospital from 1989 to 2006 to further characterize IgM+ IF in children with nephrotic syndrome. Of the 55 children with steroid-resistant or -dependent minimal change disease (MCD), 23 had IgM+ IF. Of these 23 children, 61% had microscopic hematuria at presentation, 48% (11/23) were steroid-dependent, and 48% (11/23) steroid-resistant (one underwent biopsy prior to steroid therapy). We compared the efficacy of adjuvant treatment with cyclophosphamide and cyclosporine: 18% initially treated with cyclophosphamide obtained remission, while 55% had no response; 83% obtained subsequent remission with cyclosporine. Of those initially treated with cyclosporine, 88% obtained complete or partial remission. IgM+ IF may be surrogate marker for the severity of MCD. Based on our results, children with MCD and IgM+ IF have a better response to cyclosporine than cyclophosphamide.     

Immunoglobulin synthesis by peripheral blood mononuclear cells in minimal change nephrotic syndrome

The spontaneous and pokeweed mitogen (PWM)-induced immunoglobulin synthesizing activities of circulating mononuclear cells (MNC) from minimal change nephrotic syndrome systemic (MCNS) patients in relapse (N = 13) were compared with those of patients in remission (N = 9), patients with active systemic lupus erythematosus (SLE, N = 9), and healthy controls (N = 17). Cumulative amounts of IgM, IgG, and IgA secreted over a 12-day culture period were determined in a solid phase radioimmunoassay. Mean levels of spontaneous immunoglobulin production in control cultures did not exceed 370 ng/ml. In contrast unstimulated IgM, IgG, and IgA synthesis among MCNS patients in relapse averaged 588, 1258, and 2665 ng/ml, respectively. The majority of patients exhibited synthetic activities that equalled or exceeded those of patients with active SLE. Spontaneous immunoglobulin production declined by 80 to 97% in three patients restudied in stable remission. A fourth patient with frequent relapses maintained high rates of synthesis in remission as well as in relapse. PWM stimulation increased immunoglobulin production in patients in remission and controls but failed to increase or suppressed immunoglobulin secretion in SLE patients and patients in relapse. These results suggest that MNC from MCNS patients in relapse are reversibly activated in vivo.     

Interstitial and glomerular immune cell populations in idiopathic nephrotic syndrome

Tissue samples from patients with idiopathic nephrotic syndrome [seven with minimal change (MC) and seven with mesangial proliferation (MP)] were examined for evidence of interstitial and glomerular immune cell infiltration using monoclonal antibodies to identify T cells (OKT3, TA-1), T cell subsets (OKT4, OKT8), B cells (BA-1), and monocytes and null cells (OKM1) by indirect immunofluorescence. A nuclear counterstain permitted precise enumeration of reactive and unreactive cells in comparison with five normal tissue samples, eight endstage (ES) tissue samples, and five tissue samples from patients with miscellaneous ( misc ) types of nephrotic syndrome. Interstitial cell populations in MC and MP were similar to normals except for an increase in OKM1 reactive cells in MP. ES had more numerous interstitial cells reactive with each of the monoclonal antibodies than did normal, MC, or MP. Reactive glomerular cells were most numerous in MP, intermediate in MC but only rarely observed in normal tissue samples. Most identifiable glomerular cells reacted with OKT3 or OKM1. T-cell subset populations in MC and MP glomeruli were similar. These studies reveal the presence of infiltrating immune cells in the glomeruli but not the renal interstitium of patients with idiopathic nephrotic syndrome. Whether these cells mediate proteinuria or simply mark tissue injury remains to be discerned.     

Glomerular T cell and monocyte populations in patients with IgA nephropathy

Renal tissues from 19 patients with the minimal or slight stage of IgA nephropathy were examined for evidence of glomerular T cell or monocyte infiltration using monoclonal antibodies to identify T cells (OKT3, OKT11), T cell subsets (OKT4, OKT8), and monocytes and null cells (OKM1) by indirect immunofluorescence. Renal tissues from 12 patients at the same stage of mesangial proliferative glomerulonephritis without IgA deposition were also investigated by these procedures. Light microscopic examination of the same renal tissues was also performed. Reactive glomerular mononuclear cells were found to be numerous in patients with IgA nephropathy. The most prominent type of glomerular cells was OKT8 positive. T cell subsets and OKM1 positive cells in glomeruli from patients with IgA nephropathy were significantly increased as compared to those from patients with mesangial proliferative glomerulonephritis. Focal segmental proliferation of mesangial cells was observed in glomeruli which showed an accumulation of OKT8 positive cells. It is concluded that the immuno-regulatory mechanism involving T cells and/or monocytes might be one of the exacerbating factors in patients with IgA nephropathy.     

Mononuclear cells in renal allografts. Correlation with peripheral blood T lymphocyte subpopulations and graft prognosis

Cellular infiltrates in 20 renal allograft biopsies taken at the time of acute rejection episodes were analyzed with antibodies reactive with monocytes (BRL antihuman monocyte monoclonal antibody), T lymphocyte subpopulations (OKT- and Leu-series of monoclonal antibodies), and B lymphocytes (heterologous antihuman IgM antibodies). For demonstration of the various mononuclear cells, an indirect immunoperoxidase technique was used. The number of monocytes in the infiltrates varied from 10-20%; the number of B lymphocytes was always below 10%. The T lymphocytes accounted for 50-90% of the total number of mononuclear cells. The OKT4/OKT8 ratios for the T lymphocytes in the graft infiltrates were correlated with the peripheral blood OKT4/OKT8 ratios measured by indirect fluorescence and flow cytometry. The OKT4/OKT8 ratios for perivascular infiltrates correlated far better with peripheral blood OKT4/OKT8 ratios (r = 0.72) than did the OKT4/OKT8 ratios for interstitial infiltrates (r = 0.58). Low or inverted OKT4/OKT8 ratios and low or inverted Leu 3a/Leu 2a ratios were associated with a high risk of irreversible graft rejection (P less than 0.001 for perivascular infiltrates).     

Monoclonal antibody therapy. Anti-idiotypic and non-anti-idiotypic antibodies to OKT3 arising despite intense immunosuppression

The frequency, timing, and specificity of the humoral antibody response to a murine monoclonal antibody (OKT3, IgG2a) were measured in 21 consecutive renal allograft recipients. These patients received i.v. OKT3, 1-5 mg/day for 10-20 days as treatment for acute graft rejection. Maintenance immunosuppression consisted of azathioprine and corticosteroids. Using three different assays, an antibody response was detected in 75% of the 20 patients with adequate samples. The ELISA assay of the overall IgM and IgG reactivity to OKT3 revealed that IgM anti-OKT3 appeared in 65% and IgG anti-OKT3 in 50% of the patients, reaching a peak 20-33 days after the last dose of OKT3. The IgM preceeded the IgG in most cases (P less than 0.02) and in 8 cases was detected during therapy. One patient had high levels of IgM anti-OKT3 before therapy, yet responded normally to OKT3. Interference with the therapeutic effectiveness was evident in one patient who developed IgG antibodies during therapy. His serum blocked the binding of F-OKT3 to normal lymphocytes in the presence of normal BALB/c serum. The blocking assay, done by flow cytometry, measured anti-idiotypic (Id) reactivity since the sera did not affect the binding of OKT8 (another IgG2a) or anti-Leu4 (another anti-T3), and the blocking activity remained after affinity absorption with normal mouse IgG. Using this assay, 60% of the patients made an anti-Id response. One made only anti-Id, and several had anti-Id at times when other reactivities were undetectable. Antibodies to non-idiotypic, presumably isotypic, determinants represented on OKT8 occurred in only 44%, while other reactivity (OKT4; IgG2bK) was less common (12%) and weaker. While no adverse allergic reactions occurred in this group of patients, the anti-Id antibodies, which are a prominent feature of the immune response to this and probably other monoclonal antibodies, can block their therapeutic effectiveness and can arise despite intense immunosuppression. This response may require the use of different idiotypes for prolonged or repeated courses of therapy and may be the major obstacle to the use of human monoclonal antibodies.     

A study of the T-lymphocytes and their subsets in the peripheral and spleen using OK and Leu series monoclonal antibodies--especially patients with idiopathic portal hypertension (IPH) and liver cirrhosis

The T lymphocytes and their subsets in the peripheral blood and spleen of the patients with idiopathic portal hypertension (IPH) and liver cirrhosis were analysed using OK and Leu series monoclonal antibodies. The healthy subjects showed no difference between the OK series (OKT3, OKT4, OKT8) and the Leu series (Leu3a, Leu2a). In the patients with IPH or liver cirrhosis, the percentages of OKT3 and Leu 4 positive lymphocytes (pan T), and OKT4 and Leu3a positive lymphocytes (H/I T) were similar, but the percentages of OKT8 and Leu2a positive lymphocytes (S/C T) in the patients with IPH were slightly decreased. The changes in the T lymphocyte subsets differed among the patients, but the patients with IPH and liver cirrhosis showed a slight decrease in the mean number of H/I T and a slight increase in that of S/C T after splenectomy. Pan T and H/I T in the IPH cases were similar to those of the normal spleen, but S/C T was slightly higher than that of the normal spleen. The distribution of the lymphocytes in the spleen examined by the ABC method showed that in the normal spleen, the OKT-positive lymphocytes were concentrated around the artery. The B1-positive lymphocytes were distributed on the outer portion of the OKT3-positive lymphocytes aggregate in a mantle shape. The distribution of the OKT4-positive subset and that of the OKT3-positive subset were almost the same. OKT8-staining was seen sporadically in the medullary cord, but there were slightly more OKT8-positive lymphocytes in the medullary cord in these spleens than in the normal spleen.     

Renal mononuclear inflammatory cell populations in membranous glomerulonephritis: a fine-needle aspiration biopsy study

Fine-needle aspiration biopsy (FNAB) technique was applied to the detection of alterations in renal inflammatory cell subsets in seven healthy control persons and 24 patients with membranous glomerulonephritis (MGN). Eighteen of the patients had nephrotic syndrome, and the rest had either persistent proteinuria or remission of the disease. Only one patient showed impaired renal function. Compared with the controls, FNAB revealed a significantly (p less than 0.01) increased number of mononuclear inflammatory cells (MIC, the sum in increment percentages of lymphocytes and monocytes) in MGN. This was mainly due to relative lymphocytosis; the number of monocytes was high particularly in patients with daily urine protein excretion in excess of 5 g. The duration of the disease did not clearly correlate with the degree of lympho- or monocytosis. In nine patients the results of the renal needle biopsy and FNAB could be compared. Biopsy revealed signs of MIC infiltration in 5/9 patients and FNAB in 7/9 patients. T-lymphocyte subsets in FNAB and peripheral blood were analyzed using monoclonal antibodies and indirect immunoperoxidase technique. The OKT4/OKT8 ratio in FNAB was 0.96 +/- 0.07 (s.e.m.) in patients and 1.49 +/- 0.12 in controls (p less than 0.01). In peripheral blood these ratios were 0.8 +/- 0.07 and 1.43 +/- 0.13 (p less than 0.001), respectively. The alterations were due to a decreased number of OKT4 positive lymphocytes. These studies suggest an associated interstitial accumulation of MIC and an imbalance between T helper/inducer and T suppressor/cytotoxic cells in MGN.     

Immune recovery following allogeneic bone marrow transplantation

A total of 144 evaluations of cell surface markers and cellular immune functions were carried out in 57 patients undergoing allogeneic bone marrow transplantation for acute leukemia in remission and relapse and for aplastic anemia. The periods tested were pretransplant, and 1-3, 4-6, 7-12 and more than 12 months posttransplant. The determination consisted of lymphocyte counts; lymphocyte surface marking using OKT3, OKT4, and OKT8 antibodies; and determination of adherent cells, lysozymes and antibody dependent cellular cytotoxicity (ADCC) against chicken red blood cells, human red blood cells, and CEM cells. Natural killer cells were determined against K562 target cells. Lymphoblastic responses were tested after stimulation with pokeweed mitogen (PWM), concanavalin-A (Con-A), and phytohemagglutinin (PHA). We found that the progression in the leukemic state (first remission, second remission, and relapse), prior to transplantation was paralleled by a decrease in T4 lymphocytes (976/microliter +/- 462; 411/microliter +/- 222; 372/microliter +/- 419; P = .04). There was a lack of helper cells and an inverted T4:T8 ratio beyond one year posttransplant independent of graft-versus-host disease status. Lymphocyte functions persisted to be depressed for more than one year. We found a direct correlation of T4 helper cells and an inverse correlation of T8 suppressor cells with lymphoblastic responses to mitogens. It is hoped that the longitudinal evaluations of immune functions after allogeneic bone marrow transplantation, and the characterization of the immune defects seen may lead to better immunorestorative treatments.