Quantitative bone histomorphometry and circulating T lymphocyte subsets in postmenopausal osteoporosis

To explore the influence of the immune system on the development of osteoporosis, 19 untreated postmenopausal women with osteoporosis were studied by means of quantitative histomorphometry of the ilium and an analysis of T lymphocyte subsets in the peripheral blood. Osteoporotic women had lower OKT3+ and OKT8+ counts and a higher OKT4+/OKT8+ ratio than nonosteoporotic control subjects. Linear regression analyses disclosed that the age of subjects correlated with bone mineral density (BMD; r = -0.634, p less than 0.01) and some of the histomorphometric parameters for bone formation (r = -0.694 to -0.467, p less than 0.01-0.05). The number of OKT4+ cells showed weak but significant negative correlation with the parameters for bone resorption (r = -0.549 to -0.462, p less than 0.05). In a multiple regression analysis, the advanced age, the increase in OKT3+, and the decrease in OKT4+ and OKT8+ counts were shown to be significant predictors for the decrease in BMD (R = 0.882, p less than 0.01). According to the regression formula obtained from the analysis, the parameters for bone formation were related only to the age of subjects whereas those for bone resorption were tightly associated with the number of OKT4+ and OKT8+ cells but not with the age of subjects. These results indicated that, in addition to the age factor, abnormalities of the peripheral T lymphocyte subsets, especially those of OKT4+ and OKT8+ cells, are closely associated with the decrease in bone mass in postmenopausal osteoporosis, supporting the causal relationship between T lymphocyte functions and the development of postmenopausal osteoporosis.     

Evaluation of lymphocyte subpopulations in patients with bladder cancer: a study with monoclonal antibodies

Peripheral blood lymphocyte subpopulations of 47 patients with bladder cancer were analyzed using monoclonal antibodies (OKT3, OKT4, OKT8 and OKIa7). Thirty five patients were untreated, and 12 were treated, disease-free patients. In untreated patients, the number of total lymphocytes and the percentage of OKT3+ cells decreased. Among the OKT3+ T cell population, the percentage of OKT4+ cells tended to decrease significantly, whereas the percentage of OKT8+ cells rather increased, resulting in the decrease of the OKT4+/OKT8+ ratio. These tendencies became more evident as the stage advanced. In treated, disease-free patients, no such characteristic changes were observed and the lymphocyte subpopulation apparently fell into the normal range. Furthermore, the retrospective study clearly indicated that the patients with a lower OKT4+/OKT8+ ratio showed a higher recurrence rate within a year. Analysis of peripheral blood lymphocyte subpopulations thus appears to be useful in the follow-up of bladder cancer patients, especially those with superficial tumors.     

Immunological evaluation after splenectomy for hairy cell leukemia--a case report

Immunocompetence is generally reduced in patients with hairy cell leukemia (HCL), and splenectomy is advocated by many authorities as being the initial treatment of choice for HCL. A 46-year-old Japanese man with HCL underwent a splenectomy and was studied, with respect to immunological parameters, pre- and post-operatively. Preoperative analysis of the T cell subsets showed decreases in the proportions of OKT3 positive cells, OKT4 positive cells and OKT8 positive cells; a decrease in the OKT4/OKT8 ratio; and a markedly reduced lymphocyte blastic response in peripheral blood. After splenectomy, the increase in the proportion of OKT4 positive cells was greater than in that of OKT8 cells and the lymphocyte blastic response was also improved. The patient's post-operative course has been uneventful and immunological improvement is still recognizable now, 1 year after splenectomy.     

A study of peripheral blood lymphocyte subsets in patients with bladder tumors

The analysis of peripheral blood lymphocytic subpopulations was conducted with 16 bladder tumor patients. Lymphocytes were collected centrifugally from the peripheral blood of preoperative patients with bladder tumors, and measured by means of flow cytometry. Among the stages of the bladder tumor patients, there were no clear relationships in the ratio between T cells and B cells. However, with the progression in stages a rise in OKT 8, as well as the lowering of the OKT 4/OKT 8 ratio were discerned. In relation to pTis, Leu 7 showed a higher value than that in other groups with advanced invasion. This suggests that in measurements of lymphocyte subsets in peripheral blood, it will to a certain extent be possible to estimate the preoperative immunological competence of bladder tumor patients.     

The effect of hemodialysis on lymphocyte subsets during dialysis

To clarify the effect of hemodialysis on immune response of dialysis patients, peripheral lymphocyte subsets were measured using monoclonal antibodies, OKT3, OKT4, OKT8, OKIal and Leu7. The absolute numbers of OKT3+, OKT4+, OKT8+ and OKIal+ cells were significantly lower in dialysis patients than controls. During dialysis, the relative proportions of OKT3+ and OKT4+ cells increased, and the absolute numbers of OKIal+ and OKT8+ cells decreased, 30 minutes after initiation of dialysis. At the end of the dialysis, the absolute numbers of OKT3+ and OKT4+ cells increased. The absolute number of Leu7+ cells showed low values during dialysis. These changes of lymphocyte subsets during dialysis may have influence on the immune response of dialysis patients.     

Immunological evaluation after splenectomy for hairy cell leukemia —A case Report—

Immunocompetence is generally reduced in patients with hairy cell leukemia (HCL)¹, and splenectomy is advocated by many authorities as being the initial treatment of choice for HCL.^2–4 A 46-year-old Japanese man with HCL underwent a splenectomy and was studied, with respect to immunological parameters, pre- and post-operatively. Preoperative analysis of the T cell subsets showed decreases in the proportions of OKT3 positive cells, OKT4 positive cells and OKT8 positive cells; a decrease in the OKT4/OKT8 ratio; and a markedly reduced lymphocyte blastic response in peripheral blood. After splenectomy, the increase in the proportion of OKT4 positive cells was greater than in that of OKT8 cells and the lymphocyte blastic response was also improved. The patient's post-operative course has been uneventful and immunological improvement is still recognizable now, 1 year after splenectomy.     

Lymphocyte subsets, phytohaemagglutinin responsiveness of blood lymphocytes, and interleukin 2 production in sarcoidosis.

To test the possibility that T lymphocyte subset imbalance and interleukin 2 (IL2) play a part in the impairment of cellular immune response in sarcoidosis, the proportion of T lymphocyte subsets in peripheral blood and alveolar lavage fluid from 21 patients with sarcoidosis was studied, monoclonal antibodies OKT3, OKT4, and OKT8 being used. Lectin induced production of IL2 and phytohaemagglutinin (PHA) responsiveness of peripheral blood lymphocytes were investigated. The percentage of both OKT3+ and OKT4+ T lymphocytes was significantly lower in peripheral blood from patients with sarcoidosis than in control subjects (control 63% and 46%), more so in patients with chronic sarcoidosis (44% and 23%) than in patients with recent sarcoidosis (56% and 38%). PHA induced IL2 production from peripheral blood lymphocytes did not differ between patients with sarcoidosis and control subjects. There was a significant positive correlation between PHA responsiveness and the percentage of blood OKT3+ and OKT4+ cells. Peripheral blood lymphocyte PHA responsiveness was decreased only in patients with an OKT4/OKT8 ratio of less than 1.5. Finally, late addition of exogenous IL2 to the culture medium on day 5 increased 3(H)Tdr incorporation by PHA stimulated blasts in peripheral blood lymphocytes from normal subjects, but not from those of patients with sarcoidosis. The data suggest that the impairment of cellular immune response in patients with sarcoidosis could in part reflect a decrease in the percentage of blood T helper lymphocytes and impaired IL2 receptors at the surface of stimulated lymphocytes.     

DXA测定诊断骨质疏松症的敏感指标

目的探索双能X线吸收法（DXA）骨密度测定仪测定诊断骨质疏松症（OP）的敏感指标.方法随机抽取成都地区年龄20岁以上健康男、女1456名（其中女性793名）,准确测定身高（H）、体重（W）后,采用美国Lunar公司生产DPX-L型DXA测定受试者腰椎和髋部的骨矿含量（BMC）、骨密度（BMD）.全部资料输入微机,用SPSS软件进行统计学处理.结果男、女在峰值骨量后,随年龄增加,BMC、BMC/W、BMD均逐渐下降,BMC、BMD、BMC/W累积丢失率在40岁以上组均随年龄增加而增加,女性丢失更明显（P＜0.05）,同部位、同年龄组三指标累积丢失率差异无显著性（P＞0.05）.OP检出率以BMD为参照指标最高,明显高于BMC/W、BMC二指标（P＜0.05）.结论DXA测定中BMD能较好反应BMC、BMC/W变化,是诊断OP的敏感指标.     

Gamma-interferon production capacity and T lymphocyte subpopulation after allogeneic bone marrow transplantation

T cell phenotypes after bone marrow transplantation (BMT) were investigated using monoclonal antibodies (moAbs) reactive to lymphocyte cell surface antigens. Patients' T cells showed decreased percentages of OKT4, 4A and 9.3-positive T cells, and increased percentages of OKT8, human Ia, and Leu-7-positive T cells. These changes in T cell phenotype persisted for a long period after BMT and had no correlation with the occurrence of graft-versus-host disease (GVHD). No lymphocyte activation antigens such as TIA (Tac) or transferrin receptor (5E9) were detected after BMT. The capacity of the patients' lymphocytes to produce gamma-interferon (IFN-gamma) was measured after incubation of lymphocytes with mitogen. Patients' lymphocytes produced significantly lower levels of IFN-gamma than the normal controls. This failure of IFN-gamma production showed no correlation with stimulation index of mitogen blastogenesis or changes of T cell population. Thus, not only T cell phenotype but also measurement of IFN-gamma production of lymphocyte may be useful in detecting immunological abnormalities in patients who receive BMT.     

Analysis of immune status of hemodialyzed adults: association with prior transfusions

Peripheral blood leukocytes of 29 hemodialyzed adults, 19 transfused and 10 nontransfused, were studied using immunofluorescent staining with monoclonal antibodies and in vitro measurement of natural killer (NK) cell activity. When compared with control subjects, the absolute number of leukocytes in transfused hemodialyzed patients was significantly reduced (P less than 0.01), as were the absolute numbers of OKT11+ cells (P less than 0.01), and OKT4+ cells (P less than 0.0001). The percent representation of OKT11+ and OKT4+ cells was also significantly lower among transfused hemodialyzed patients (P less than 0.01 and 0.001, respectively), and this loss of OKT4+ cells resulted in a decrease in the ratio of OKT4+/OKT8+ cells (P less than 0.01). The absolute number of Leu-7+ cells was also decreased in the transfused group (P less than 0.05). A decrease in in vitro NK cell activity was present in both transfused and nontransfused hemodialyzed subjects. Whether these differences in peripheral blood lymphocytes were induced by the erythrocyte transfusions could not be determined; however, if they reflect changes in central lymphoid tissues, then these results may help explain the prolonged survival of renal allografts in transfused individuals.     

Changes in the immune response in postoperative patients while running the ventricular assist device (VAD)

We examined the immunological parameters in two patients who were assisted with LVAD in postoperative LOS. Both patients demonstrated transient consumption of complements immediately after running the LVAD. The serum concentrations of immunoglobulins (IgG, IgM, IgA) showed no significant decrease. Both patients had significant postoperative lymphopenia, but recovered in a week. A decrease in the number of T cells was observed in both patients. The OKT4/OKT8 ratio initially increased in one patient, but gradually declined postoperatively. In the other patient, the OKT4/OKT8 ratio decreased. The lymphocyte response to mitogen (PHA) remained significantly depressed when the number of lymphocytes returned to the normal level, suggesting that the lymphocytes were functionally impaired. Such findings are generally observed after cardiac surgery. Thus, LVAD does not affect the immune response.     

混合培养的供、受者骨髓细胞过继回输延长小鼠移植心脏存活时间

目的 将供、受者骨髓细胞经混合培养后过继回输,以观察其对同种异体移植心脏存活时间和受者免疫功能的影响.方法 取Balb/c小鼠和C57BL/6J小鼠的骨髓细胞,进行混合培养.配制含Balb/c小鼠和C57BL/6J小鼠脾淋巴细胞的混合淋巴细胞反应体系(MLR)以及含Balb/c小鼠和C3H小鼠脾淋巴细胞的MLR,分别加入混合培养的骨髓细胞,观察其对MLR中细胞增殖的影响.以C57BL/6J小鼠为供者,Balb/c小鼠为受者行腹腔异位心脏移植,实验分为4组:(1)移植对照组,受者仅进行心脏移植,不作其他处理;(2)实验对照组,心脏移植后给予西罗莫司灌胃;(3)实验组,移植手术结束前注射混合培养的骨髓细胞1×10~7个,术后给予西罗莫司;(4)第三方对照组,受者接受C3H小鼠的移植心脏,手术结束前注射混合培养的骨髓细胞1×10~7个,术后给予西罗莫司.记录移植心脏存活时间;移植心脏停跳当日,取受者外周血,检测CD4~+ CD25~+ T淋巴细胞的比例及供者来源的H-2K~b细胞的比例.结果 加入混合培养的骨髓细胞后,Balb/c和C57BL/6J的MLR的淋巴细胞增殖率低于Balb/c和C3H的MLR.实验组移植心脏的存活时间长于其他3组(P<0.05).实验组CD4~+CD25~+T淋巴细胞的百分率高于其他3组(P<0.05).实验组外周血中H-2K~b细胞的比例高于其他3组(P<0.05).结论 受者输注混合培养的供、受者骨髓细胞可在一定程度上调节免疫应答,延长小鼠移植心脏的存活时间,该作用具有供者抗原特异性.     

Immunological and clinical observations in diabetic kidney graft recipients pretreated with total-lymphoid irradiation

In a feasibility study, twenty patients with end-stage diabetic nephropathy were treated with fractionated total-lymphoid irradiation (TLI, mean dose 25 Gy), before transplantation of a first cadaveric kidney. During radiotherapy, only one patient had a serious side effect (bone marrow depression). After transplantation four patients died (one of a myocardial infarction, one of ketoacidosis, and two of infections occurring during treatment of rejection crises). One graft was lost because of chronic rejection. The other 15 patients have a functioning graft (mean follow-up 24 months) and receive low-dose prednisone alone (less than 10 mg/day, n = 11) or in conjunction with cyclosporine (n = 4) as maintenance immunosuppressive therapy. A favorable clinical outcome after TLI (no, or only one, steroid-sensitive rejection crisis) was significantly correlated with a high pre-TLI helper/suppressor lymphocyte ratio, a short interval between TLI and the time of transplantation, and the occurrence of functional suppressor cells early after TLI. The most striking immunological changes provoked by TLI consisted of a long-term depression of the mixed lymphocyte reaction and of the phytohemagglutinin, and Concanavalin A or pokeweed-mitogen-induced blastogenesis. A rapid and complete recovery of the natural killer cell activity was observed after TLI. A permanent inversion of the OKT4+ (T helper/inducer) over OKT8+ (T suppressor/cytotoxic) lymphocyte ratio was provoked by a decrease of the OTK4+ subpopulation, together with a supranormal recovery of the OKT8+ lymphocytes. A majority of the latter lymphocytes did also express the Leu 7 and the Leu 15 phenotype.     

Long-bone changes after pamidronate discontinuation in children and adolescents with osteogenesis imperfecta

Cyclical intravenous pamidronate is a widely used symptomatic therapy in moderate to severe osteogenesis imperfecta (OI). The effects of treatment discontinuation on long bone development have not been characterized. In this observational study we used peripheral quantitative computed tomography to assess the radius at the distal metaphysis and at the diaphysis in 23 young OI patients (11 female) who had received pamidronate for at least 3 years. Measurements were performed twice, at the time of treatment discontinuation (when the age of the patients ranged from 5.9 to 21.3 years) and at an average of 1.9 years (range 1.5 to 2.4 years) later. At the time of pamidronate discontinuation, all but one of the patients who were below 15 years of age (n=14) had a positive age- and sex-specific z-score for bone mineral content (BMC) at the metaphysis, resulting in a mean z-score of +2.0 (SD=1.0) for this subgroup. In contrast, patients aged 15 years or older (n=9) had an average metaphyseal BMC z-score of -1.5 (SD=1.5). After pamidronate discontinuation, metaphyseal BMC z-score decreased by an average of 2.4 (SD=2.0) in the whole group. The change in BMC z-score was growth-dependent, as BMC z-scores decreased by about 2 or more in all patients in whom distal radius growth plates were open when pamidronate was discontinued. In contrast, none of the 11 patients with closed distal radius growth plates experienced a decrease in metaphyseal BMC z-score by more than 2. At the diaphysis, the average BMC z-score was low at the time of the last pamidronate infusion [z-score -1.7 (SD=1.4)]. After pamidronate discontinuation, the average diaphyseal BMC z-score decreased by only 0.3 (SD=0.4). In summary, this study shows that the effect of pamidronate discontinuation is much larger at the radial metaphysis than at the diaphysis and is dependent on growth. Metaphyseal bone tissue added by longitudinal growth after treatment discontinuation has a lower density than tissue created during treatment. It is possible that this produces zones of localized bone fragility after pamidronate treatment is stopped in growing children.     

体重对成年女性骨矿含量及其标化的影响研究

目的分析不同体重组人群[小体重组(<45 kg)、标准体重组(45~60 kg)、大体重组(>60 kg)]的骨矿含量结果,探索体重对骨矿含量及其标化的影响。方法纳入290例50~80岁绝经后女性,按体重大小分为大体重组、标准体重组、小体重组。通对不同体重组人群的腰椎L_1~4、股骨的骨矿含量及骨密度进行测量,并对测量结果进行比较分析。同时将年龄、体重作为应变量,腰椎或股骨颈骨矿含量作为自变量,进行多重线性回归分析。结果低体重组的腰椎或股骨颈骨密度T值、骨矿含量均明显低于标准体重组和高体重组人群。随着年龄增长,L_1~4及股骨颈骨矿含量均下降,年龄每升高1岁,L_1~4骨矿含量下降0.364 g、股骨颈骨矿含量下降0.031 g;随着体重增长,L_1~4及股骨颈骨矿含量均升高,体重每增长1 kg,L_1~4骨矿含量升高0.548 g、股骨颈骨矿含量升高0.025 g。结论成年女性的体重与骨矿含量显著正相关,因此,体重是骨矿含量标化的重要指标之一,可以避免骨质疏松的漏误诊。     

Sex-specific relationships between insulin resistance and bone mineral content in Korean adolescents

The aim of this study was to examine sex-specific relationships between insulin resistance (IR) and bone mineral content (BMC) according to age group and weight status. A population-based sample of 618 Korean adolescents (315 male and 303 female), aged 10–19 years from the Fourth Korea National Health and Nutrition Examination Survey, 2009. They were divided into three age groups (10–12; 13–16; 17–19 years) and two weight groups (non-overweight vs. overweight). IR was assessed using the homeostatic model of assessment of IR (HOMA-IR). Soft tissue composition (fat and lean mass) and BMC of the whole body, proximal femur, and lumbar spine were measured by dual energy X-ray absorptiometry. Adiposity (body mass index, waist circumference, or soft tissue composition), age, height, total cholesterol, triglycerides, high density lipoprotein cholesterol, alkaline phosphatase, serum vitamin D, dietary calcium and energy intake, and menarche for females were adjusted using general linear models of a complex sampling design. Higher HOMA-IR was associated with a decrease in BMC in male adolescents aged 13–19 years after adjustment for adiposity and other confounders, while the associations were not significant in male adolescents aged 10–12 years and female adolescents. After adjustment for adiposity and confounders, the inverse associations between HOMA-IR and BMC were more consistent in non-overweight male adolescents than in other weight groups (overweight males and non-overweight and overweight females). The unfavorable effect of IR on BMC appears to be more obvious in males aged 13–19 years or non-overweight males than in females.     

A randomized study on the effects of estrogen/gestagen or high dose oral calcium on trabecular bone remodeling in postmenopausal osteoporosis

Thirty-seven patients with postmenopausal crush fracture osteoporosis were randomized to oral cyclic estrogen/gestagen (n = 20) or oral calcium (2000 mg elemental calcium per day) (n = 17). Fourteen in each group completed 1 year of treatment. Iliac crest bone biopsies were obtained after intravital double labeling with tetracycline before and after treatment in 10 patients on estrogen/gestagen and 11 patients on calcium. In the estrogen/gestagen group the activation frequency in trabecular bone decreased from 0.52 + 0.11 (SEM) year-1 to 0.27 + 0.08 year-1 (p less than 0.01). No significant changes were found in resorption or formation periods. The osteoid surfaces and the mineralizing surfaces decreased (p less than 0.05), whereas the decrease in eroded surfaces was insignificant. Furthermore, no significant changes were observed in final resorption depth, wall thickness or bone balance per remodeling cycle. Serum alkaline phosphatase and renal hydroxyproline excretion decreased during treatment (p less than 0.002), whereas the lumbar bone mineral content (BMC) increased (p less than 0.01). In the calcium group the extent and thickness of osteoid surfaces decreased (p less than 0.05) without significant changes in activation frequency. Serum alkaline phosphatase and renal hydroxyproline excretion decreased during treatment (p less than 0.02). No significant changes were observed in lumbar BMC or the other histomorphometric parameters. The study supports that the positive effect of estrogen/gestagen on BMC can be explained by a reduction in the activation frequency of new remodeling cycles leading to a decreased remodeling space and an increase in mean bone age. There is no evidence of a positive balance per remodeling cycle.(ABSTRACT TRUNCATED AT 250 WORDS)     

Evaluation of bone mineral density in postmenopausal women in Kuwait

Menopause is the major risk factor for the loss of bone mineral density (BMD) and bone mineral content (BMC) in women. In this study, we determined the prevalence of osteoporosis in postmenopausal women in Kuwait and compared it with that of other Middle East and west countries. Two thousand two hundred ninety-six postmenopausal women ranging in age from 40 to 87yr were included in the study and divided into 4 age groups by decade. We measured body weight, height, body mass index (BMI), BMD, and BMC. The mean age, height, and weight were 59.1+7.9yr, 154.7+6.5cm, and 77.3+14.9kg, respectively. The mean BMI and BMC were 32.4+6.6kg/m(2) and 0.9+0.14g/cm(2), respectively. The average T-scores for the hip and lumbar spine were -0.280+1.2 and -1.297+1.33, respectively. BMC significantly decreased with age from 0.95 to 0.81g/cm(2). Four hundred forty-four (19.3%) were found to have osteoporosis. The incidence of osteoporosis significantly increased from 4.3% to 39.9% with age, which is lower than that reported for Saudi (40%) and Moroccan women (39.6%) and higher than that for US/European (31%) and Lebanese women (11%).     

Immunohistochemical study using a double fluorescent staining on the regional lymph nodes of gastric cancers]

For the purpose to investigate the immunological anti-tumor function of lymphocytes in the regional lymph nodes of gastric cancer, a double staining procedure using several monoclonal antibodies against the surface membranes of T and NK cells with fluorescent antibody technique was conducted. The number of CD11b+ cells out of CD8+ cells was small with almost all being CD11b- belonging to cytotoxic T cells. Leu8+ cells and Leu8- cells out of CD4+ cells were recognized in equal numbers and were reciprocal. The ratio of CD4+ Leu8-/CD4+ demonstrated an increase in the group injected with OK-432. The ratio of OKT9+ or OKIa1+ occupied in CD8+ or CD4+ cells was only several percent, and few in number. An increase for the ratio of CD4+ OKT9+/CD4+ was observed in the group injected with IL-2. Similar increases for the ratio of CD4+ OKIa1+/CD4+ were obtained in the group injected with OK-432 and in the metastatic group, respectively. The Leu7+ CD16+ cells were not observed. The Leu7- CD16+ cells were observed in a part where metastases were focused. These results indicated that lymphocytes in regional lymphocyte of gastric cancer might hold the hidden anti-tumor effect, but did not display the full function without preoperative intratumor injection of BRM such as IL-2 or OK-432.     

Interest in and limitations of monoclonal anti-T-cell antibodies for the follow-up of renal transplant patients

The OKT series of anti-T-cell monoclonals has been used on 442 occasions in 41 renal allograft recipients in a 6-12 month follow-up study. Standard immunosuppressive therapy (including antithymocyte globulin in 26 patients) tended to decrease the helper-inducer/suppressor-cytotoxic cell ratio (OKT4/OKT8). Conversely, 71% of 35 renal failure episodes were associated with increased OKT4/OKT8 ratios. Twenty-three percent of renal failure episodes were associated with dramatically decreased OKT4/OKT8 ratios. At least half of these cases could be explained by a cytomegalovirus infection. In fact, similar infections were found in 6 out of 17 patients with low OKT4/OKT8 values in the absence of renal failure. These results prompt us to use anti-T cell monoclonals for the diagnosis of rejection because only nine episodes of transient increase in the OKT4/OKT8 ratio were observed in the absence of rejection. The interest of this new method for the immunological follow-up of transplanted patients is, however, limited by the difficulty in interpreting a significant percentage of tests because of (1) the presence of doubly labeled cells (OKT4+OKT8+) or the significant discrepancy between the number of OKT3+ cells and total cells labeled with OKT4 and/or OKT8 antibodies; (2) gross lymphocytopenia--most often observed in patients receiving antithymocyte globulins plus steroids; and (3) the clinically unexplained shifts in the T cell subset ratios mentioned above.