Apoptosis and oxidative status in peripheral blood mononuclear cells of diabetic patients

We have compared the concentrations of intracellular glutathione (GSH), glutathione-dependent antioxidative enzymes, the cell death rate and immunophenotype profile of peripheral blood mononuclear cells (PBMC) from healthy donors and from patients with insulin-dependent type I (IDDM) or non insulin-dependent type II (NIDDM) diabetes mellitus. The IDDM and NIDDM patients had above-normal absolute lymphocyte counts, whereas the percentages of CD3, CD4 and CD8 T lymphocytes were significantly reduced. In contrast, the absolute number and percentage of B lymphocytes was higher in diabetic patients than in healthy donors. The low intracellular reduced glutathione (GSH) and the unbalanced profile of key enzymes involved in GSH metabolism, gamma glutamyltransferase (-GT) and glutathione-S-transferase (GST), account for the increased oxidative status of PBMC from diabetic patients. The plasma membranes of PBMC from diabetic patients were less permeable to propidium iodide than those of PBMC from healthy donors, indicating that the apoptotic cell death rate was lower in the cells from diabetic patients. These differences are potentially useful markers of pathogenic metabolic changes which occur during clinical diabetes and if they are confirmed could be used to identify the onset of diabetes.     

In vivo expression of perforin by CD8+ lymphocytes in autoimmune disease. Studies on spontaneous and adoptively transferred diabetes in nonobese diabetic mice

A potent cytolytic pore-forming protein (perforin or cytolysin) has previously been found to be associated with the cytoplasmic granules of CTL and NK cells. Inasmuch as all previous studies on perforin have been conducted with cultured CTL and NK cell lines, it is not clear whether perforin may play a role in the cytotoxicity mediated by CTL that have been primed in vivo. In this study, we investigated the presence of perforin in pancreata from nonobese diabetic (NOD) mice, which have been studied as a model of autoimmune, insulin-dependent (type I) diabetes mellitus. Whereas adult NOD mice spontaneously develop diabetes, it is possible to induce diabetes in young, irradiated NOD mice by adoptive transfer of splenocytes obtained from diabetic donors. By means of immunohistochemical analysis, we were able to detect perforin Ag in a small subpopulation of CD8+/Thy-1+/asialo GM1-/CD4- lymphocytes in the pancreatic islets of animals undergoing both spontaneous and adoptive transfer-mediated insulitis. Perforin+/CD8+ lymphocytes were found in small clusters and were observed to display the morphology of large granular lymphocytes. These observations show for the first time the presence of perforin-containing CD8+ lymphocytes in tissues of animals undergoing autoimmune disease.     

Natural killer cells in patients with pulmonary tuberculosis]

In the existent literature the number of works devoted to the subpopulation of natural killer cells (NK) is not significant. The purpose of the present study was to determine the NK content in the blood of pulmonary tuberculosis patients; to establish correlation of the level of their content with the content of the previously studied T-lymphocyte subpopulations; to determine the intensity of the fluorescence of NK, CD3+, CD4+, CD8+ lymphocytes. The data were obtained on the significant increase in the NK mean level in tuberculosis patients (20.37 +/- 1.74) as compared with that in healthy subjects (12.77 +/- 2.56). The NK fluorescence intensity (56.33 +/- 2.28) conditioned by the Fc-receptor expression intensity is significantly lower than the analogous index in healthy volunteers (82.4 +/- 7.69). The NK level in the blood of tuberculosis patients correlates with the content of CD3+, CD8+ lymphocytes as well as with the CD4+/CD8+ index.     

Inhibition of autoimmune diabetes by Fas ligand: the paradox is solved

Previous reports that diabetogenic lymphocytes did not induce diabetes in nonobese diabetic (NOD)-lpr mice suggested the critical role of Fas-Fas ligand (FasL) interaction in pancreatic beta cell apoptosis. However, recent works demonstrated that FasL is not an effector molecule in islet beta cell death. We addressed why diabetes cannot be transferred to NOD-lpr mice despite the nonessential role of Fas in beta cell apoptosis. Lymphocytes from NOD-lpr mice were constitutively expressing FasL. A decrease in the number of FasL+ lymphocytes by neonatal thymectomy facilitated the development of insulitis. Cotransfer of FasL+ lymphocytes from NOD-lpr mice completely abrogated diabetes after adoptive transfer of lymphocytes from diabetic NOD mice. The inhibition of diabetes by cotransferred lymphocytes was reversed by anti-FasL Ab, indicating that FasL on abnormal lymphocytes from NOD-lpr mice was responsible for the inhibition of diabetes transfer. Pretreatment of lymphocytes with soluble FasL (sFasL) also inhibited diabetes transfer. sFasL treatment decreased the number of CD4+CD45RBlow cells and increased the number of propidium iodide-stained cells among CD4+CD45RBlow cells, suggesting that sFasL induces apoptosis on CD4+CD45RBlow "memory" cells. These results resolve the paradox between previous findings and suggest a new role for FasL in the treatment of autoimmune disorders. Our data also suggest that sFasL is involved in the deletion of potentially hazardous peripheral "memory" cells, contrary to previous reports that Fas on unmanipulated peripheral lymphocytes is nonfunctional.     

麻醉剂七氟烷与地氟烷对患者中性粒细胞和T细胞的影响研究

目的:探讨腹腔手术过程中七氟烷与地氟烷麻醉对患者中性粒细胞和T细胞的影响。方法:随机选取85例年龄在20-60岁的患者,将患者随机分成二组,分别使用七氟烷与地氟烷进行麻醉。在麻醉前30 min和麻醉后2-24 h分别检测患者白细胞、淋巴细胞、中性粒细胞的数量、T辅助淋巴细胞(CD4)、细胞毒性T淋巴细胞(CD8)、自然杀伤性淋巴细胞、激活T淋巴细胞的百分比,CD4/CD8的比值,血浆皮质醇含量。结果:二组患者白细胞与中性粒细胞麻醉24 h后显著升高(P0.01)。地氟烷组中性粒细胞数量显著高于七氟烷组(P0.05)。CD4细胞的百分比在七氟烷组中无显著改变,在麻醉后2h,地氟烷组(31.93%[6.65%])同基础值(36.41%[6.92%])相比显著降低(P0.05),24 h后进一步下降(30.52%[8.01%])(P0.01)。七氟烷组CD4/CD8比值一直无变化,但地氟烷组在麻醉2 h降低(1.51[0.52]),24 h明显升高(1.82[0.72])(P0.05)。结论:与地氟烷相比,七氟烷对中性粒细胞群与T细胞群的影响较小。     

Interrelation between the activity of hepatitis, liver fibrosis and immune status in children with chronic hepatitis B and C

The lesion of the liver in viral hepatitis was found to depend on the state of the immune system. Relationship between the content of lymphocyte subpopulations (CD3+, CD4+, CD8+, CD20+) in the blood and immunoglobulins (IgG, IgM, IgA) with parameters of semi-quantitative evaluation of the activity of hepatitis and the stage of liver fibrosis in children with chronic virus hepatitis B, C, B + C was studied. The characteristic feature of all hepatitis was a decrease in the number of T lymphocytes CD4+ below the normal level and an increase in the content of B lymphocytes. The correlation between the morphological activity of hepatitis and the amount of T lymphocytes CD8+ was established only in chronic hepatitis B. In chronic hepatitis B and B + C the absolute amount of blood lymphocytes decreased with the increase of the age of the patients, but in chronic hepatitis B this was accompanied by the decrease of the morphological activity of hepatitis and in hepatitis B + C by its increase. The amount of lymphocytes CD4+ rose with the increase of liver fibrosis in chronic hepatitis B. In children with chronic hepatitis C and B + C the amount of blood lymphocytes was found to be unrelated to the morphological activity of hepatitis.     

Quantitative assay of CD8+ (cytotoxic and suppressor) lymphocytes in patients with both asymptomatic and symptomatic HIV infection]

Lymphocytes CD8+ have been assayed prospectively in 245 individuals infected with HIV. Percentage and number of CD8+ have been nearly two-fold higher in asymptomatic patients or patients with lymphadenopathy than those in the control group. The number of CD8+ lymphocytes has been rapidly decreasing parallel to the progression of HIV (ARC and AIDS), while their percentage has increased--however insignificantly. There has been a positive correlation between the number of CD4+ and CD8+ cells and all clinical stages of HIV infection.     

Distribution of peripheral blood leukocytes in acute heatstroke.

We examined 11 heatstroke patients (mean rectal temperature 41.4 +/- 0.3 degrees C) and 40 healthy subjects to determine the effects of hyperthermia on peripheral blood leukocyte distribution. Precooling samples were taken on admission. Whole blood was incubated with conjugated monoclonal antibodies, and erythrocytes were eliminated by FACS lysing solution. Lymphocyte subsets were detected by specific mouse monoclonal antibodies: Leu-4/CD3+ (T-cells), Leu-3a/CD4+ (T-helper cells), Leu-2a/CD8+ (T-suppressor-cytotoxic cells), Leu-11/19/CD16+/CD56+ (natural killer cells), and Leu-12/CD19+ (B-cells). Immunofluorescence was measured with a flow cytometer. The number of circulating leukocytes and lymphocytes was significantly increased in heatstroke patients. This lymphocytosis was mainly due to an increase in T-suppressor-cytotoxic cells and natural killer cells. The absolute number of lymphocytes and T-suppressor-cytotoxic cells significantly correlated with the degree of hyperthermia (r = 0.62, P = 0.04; r = 0.751, P = 0.007, respectively). There was a significant decrease in the percentages of T-, B-, and T-helper cells and increase in T-suppressor-cytotoxic and natural killer cells, giving a marked decrease in the ratio of T-helper to T-suppressor-cytotoxic cells. We conclude that heatstroke is associated with leukocytosis and significant alteration in absolute number and percentage of circulating lymphocyte subpopulations.     

Anti-interferon activity of peripheral blood leukocytes in patients infected with HIV-1

The study was aimed at anti-interferon activity of different components of the in vitro system cells MT-4--HIV-1 as well as the culture fluid of peripheral blood leukocytes from patients at different stages of HIV infection and from patients with mixed infection (HIV and chronic hepatitis C) in comparison with patients infected with hepatitis C virus alone and healthy persons. Anti-interferon activity was detected in all groups of patients, its detection rate varying within 33% and 68%. The tendency towards increased detection rate of anti-interferon activity in HIV-infected patients in parallel with decreased number of CD4+ lymphocytes was noted. These data made it possible to suggest that increased detection rate of anti-interferon activity in the culture fluid of peripheral blood leukocytes from HIV-infected patients in parallel with decreased number of CD4+ lymphocytes could result from pathogenetic processes in the body, leading to a decrease in therapy effectiveness of HIV-infected patients with the preparations of alpha-interferon, especially in patients with a low content of CD4+ cells.     

The role of monocytes/macrophages in TCR-zeta chain downregulation and apoptosis of T lymphocytes in malignant pleural effusions

The aim of this study was to assess alterations of zeta chain expression and their relation to apoptosis of T lymphocytes. T lymphocytes were obtained from malignant pleural effusions (MPE) of 15 patients. The work focused on TCR-zeta chain expression, apoptosis of T lymphocytes, and on the content of monocyte/macrophages in effusions. Analysis was performed using three color flow cytometry combining CD3, TCR-zeta and TUNEL reaction. The content of tumor and monocyte/macrophage cells has been determined using CD3, CD14, and cytokeratins, as markers of distinct cell subpopulations. Our findings strongly indicate that decreased zeta chain expression in T cells depends on the content of monocyte/macrophage cells, and that the range of the decrease is inversely proportional to the number of monocytes/macrophages in the effusion. Those having low zeta chain expression were the main subpopulation of T cells undergoing apoptosis. These data suggest that decreased zeta chain expression in T cells in MPE may be related to the abundance of monocyte/macrophages in effusions.     

Immunohistochemical characterization of the cellular infiltrate in Jorge Lobo's disease

Few studies have been conducted to evaluate the cellular composition of the granulomatous lesions induced by Lacazia loboi. Thus, the objective of the present study was to characterize the mononuclear cell population present in cutaneous lesions obtained from 15 patients with Jorge Lobo's disease. Histological sections were stained with hematoxylin-eosin and methenamine silver and the following mononuclear cells were identified by immunohistochemistry: T lymphocytes (CD3+), helper T lymphocytes (CD4+), cytotoxic T lymphocytes (CD8+), B lymphocytes (CD20+), plasma cells (CD79+), natural killer cells (CD57+) and histiocytes (CD68+). This study showed that the inflammatory infiltrate mainly consists of histiocytes and multinucleated giant cells, in addition to the presence of a large number of fungal cells. The identified inflammatory cells showed the following frequency: CD68+ histiocytes > CD3+ T lymphocytes > CD4+ T > CD8+ T lymphocytes > CD57+ natural killer cells > CD79+ plasma cells > CD20+ B lymphocytes. Based on the findings of a large number of fungal cells in the infected tissues and the disorganized cell arrangement in the granuloma, we hypothesize that patients with Jorge Lobo's disease present immunoregulatory disturbances, which are likely to be specific and perhaps responsible for the lack of containment of the pathogen.     

An epidemiological analysis of monitoring of the immune status of chernobyl nuclear accident liquidators for early detection of risk groups and diagnosis of cancer diseases. Communication 2. Dependence of the rate and changes in the immune status on radiation accident risk factors

It was demonstrated that malignant neoplasms (MNs) most frequently develop in the liquidators involved in the cleanup activities after the Chernobyl nuclear accident (ChNA) in 1986 (43.75%) and the liquidators involved in long-term activities, one-quarter of whom also participated in the cleanup in 1986. The specific features of the immune status (IS) depending on the period and year of the cleanup activities in the ChNA area were determined. In the cohort with a confirmed MN diagnosis, the observed deviations in the people who participated in these activities only in 1986 and on a permanent basis, including 1986, coincided in their pattern but differed in the magnitude of changes in immunological characteristics. Long-term participation during the extreme period and, correspondingly, higher exposure caused an increase in the contents of CD8⁺ T cells, CD16⁺ lymphocytes, and activated T lymphocytes, as well as decreases in the total immunoregulatory index, percentage of CD3-16/56⁺ (NK), and total IgE and a more pronounced deficiency in B lymphocytes. Differences between the groups of liquidators who participated in the cleanup activities in 1986 and 1987 were detected. The liquidators-1987 display the most pronounced deviations in the IS characteristics. Permanent presence in the ChNA area in 1986 and 1987 caused a similar effect but the magnitude of changes in the content of CD4⁺ T lymphocytes (↓), content of CD8⁺ T lymphocytes (↑), and immunoregulatory index (↓) was considerably higher in liquidators-1987. The permanent cleanup activities in Chernobyl in 1987 caused a deficiency in CD4⁺ T lymphocytes; more pronounced increase in the content of CD8⁺ T lymphocytes; decrease in CD4⁺/CD8⁺ index; change in the ratio of NK-T to NK cells; elevation in the counts of CD95⁺, HLA-DR⁺, and activated T lymphocytes; and decrease in the level of total IgE. Any increase in the expression of cell activation markers was undetectable in the liquidators-1986 involved in the cleanup activities on a long-term basis. The specific features of the IS changes depending on the dose of external γ-irradiation were determined. It was shown that the MN rate in liquidators increases with age relative to the number of examined people in each age cohort. The differences in the IS age-related dynamics in liquidators with and without MNs were detectable in stable contents of CD3⁺, CD4⁺, and CD8⁺ T lymphocytes; immunoregulatory index; and contents of CD95⁺ cells and serum IgA at ages of 40 to 70 years old with a subsequent decrease in these parameters and elevation in the content of CD8⁺ T lymphocytes with age in the absence of MNs; continuous increase in CD3-16/56⁺ (NK) cells in the presence of MNs; and decreases in these values after 70 years in the absence of MNs. In both groups of liquidators older than 70 years (with and without MNs), a deficiency in the T-cell component (CD3⁺ and CD4⁺ T lymphocytes and CD4⁺/CD8⁺ ratio) and an increase in the counts of CD8⁺ T lymphocytes are characteristic of the IS. A growing deficiency in CD4⁺ T lymphocytes during the monitoring on the background of increasing content of CD8⁺ T lymphocytes, which additionally contributes to the weakening of immune regulation due to progressing abnormalities in the ratio of regulatory T-lymphocyte subpopulations, can be regarded as one of the characteristics that suggest an adverse life expectancy prognosis for people with MNs.     

Cellular composition of spleen and peritoneal exudate in mice after injection of cyclophosphamide-treated L 1210 leukemia cells

The composition of peritoneal exudate and spleen cells of CD2F1 mice after fourfold i.p. administration of L 1210 leukemia cells treated with cyclophosphamide (L 1210-CY cells) were examined. The number of cells in peritoneal cavity did not increase, however, the spleen weight rose after administration of L 1210-CY cells. The per cent of lymphocytes T was increased 2.5 times but the content of macrophages and lymphocytes B was normal in the peritoneal cavity after L 1210-CY cells injections. In the spleen an 1.4 times increase of the per cent of lymphocytes B, but normal level of macrophages and lymphocytes T were observed.     

Effect of chronic psychoemotional stress on subpopulation spectrum of T-lymphocytes in immunocompetent organs in male mice

Subpopulation spectrum of T lymphocytes (CD3+, CD4+, CD8+, CD25+, and CD3(+)25+) in thymus, spleen and inguinal lymphatic nodes have been studied in male mice after 20 days of psychoemotional stress produced by social defeats in daily agonistic confrontations. A reduction of total number of cells, of absolute numbers of all researched subpopulations of lymphocytes and % CD3+ cells in thymus of submissive mice was shown in comparison with intact animals. Reduction of total number of splenocytes and absolute numbers of CD4+ and CD8+ lymphocytes has been observed in a spleen of submissive mice. Besides, % CD3+, CD25+, CD4+ and CD25+ cells were increased in these animals in comparison with intact mice. The absolute number of cells with CD8 phenotype was increased in inguinal lymphatic nodes. The data obtained suggest that the chronic psychoemotional stress is accompanied by serious changes of the cellular link of immunity. The effect of chronic emotional social stress on mutual interaction of the central and peripheral links of immunity has been discussed.     

Participation of the CD27 antigen in the regulation of IL-2-activated human natural killer cells.

The CD27 Ag is expressed by the majority of resting T lymphocytes and appears to play a crucial role in T cell activation. We found that some resting peripheral blood NK cells also express CD27. Furthermore, CD27 expression was up-regulated on NK cells stimulated by IL-2. The cytolytic activity of IL-2-activated, but not resting, NK cells was inhibited by an anti-CD27 mAb (anti-1A4). However, anti-1A4 did not affect conjugate formation between IL-2-activated NK cells and tumor cell targets. In contrast, anti-1A4 inhibited CD2-mediated calcium mobilization and the serine esterase activity of NK cell granules. These inhibitory effects could be mediated in part by increase in intracellular cAMP levels induced by anti-1A4. Our results suggest that the CD27 Ag plays an important role in the regulation of activated NK cells.     

HLA-DR expression on CD8 lymphocytes from gastric mucosa in urease-positive and urease-negative gastritis

Abstract We isolated lymphocytes from chronically inflamed gastric mucosa. We analysed the expression of IL-2 receptors (CD25), transferin receptors (CD71) and HLA-DR molecules on T lymphocytes by flow cytometric analysis in 16 patients with urease-positive and in 7 patients with urease-negative chronic gastritis. In G0, G1 and G2 histological type (Sydney classification) of gastritis the number of lymphocytes obtained from the gastric mucosa biopsies was too low for the flow cytometric analysis. However, in G3 histological type of chronic gastritis we obtained enough cells for the flow cytometric analysis in 75 %. We demonstrated a significant increase in HLA-DR expression on CD8 cells from patients with urease-positive gastritis compared to urease-negative gastritis. We also observed a statistically non-significant increase in HLA-DR expression on CD3 cells, and in CD71 expression on both CD3 and CD8 cells in urease-positive gastritis. However, no difference in CD25 expression was found between the two types of gastritis.     

Reactivity of lymphocytes to a progesterone receptor-specific monoclonal antibody

In this study we present evidence for reactivity of pregnancy lymphocytes, but not nonpregnancy lymphocytes, with the progesterone receptor-specific monoclonal antibody mPRI. Using an avidin-biotin peroxidase detection system, we found a nuclear staining in 14.6 +/- 3.7% (mean +/- SEM, N = 27) of pregnancy lymphocytes, while only 0.47 +/- 0.33% (mean +/- SEM, N = 15) of nonpregnancy lymphocytes reacted with the antibody. To characterize the receptor-bearing subset, CD8+ and CD4+ cells were depleted by complement-dependent lysis. Depletion of CD8+ cells was accompanied by 62 +/- 18% loss of progesterone receptor-bearing cells, while depletion of CD4+ cells resulted in a twofold increase in the number of positively staining lymphocytes. In nonpregnancy lymphocytes a 3-day PHA treatment, as well as allogeneic stimulation, resulted in a significant increase in the number of receptor-containing cells. These results suggest that pregnancy, but not nonpregnancy, lymphocytes contain progesterone binding structures, and that these are inducible by mitogenic or alloantigenic stimuli.     

The surface phenotype and enzymatic cytochemical markers of B cells during pokeweed mitogen activation

Methods of monoclonal antibodies, cytochemistry and rosette formation have been used to study antigens, receptors and enzymatic activity of peripheral blood lymphocytes during pokeweed mitogen-stimulation. After 18-20 hours in mitogen-stimulated cultures there is a decrease in number of T-lymphocytes that express CD7, CD5, CD4, CD8 antigens and of E-receptors and cells with the local activities of acid phosphatase and acid non-specific esterase. The number of lymphocytes with E37-, FcM- and M-receptors and of cells with granular PAS-reaction increased. Blast cells were revealed after 40-48 hours. Approximately 50% blast cells forming E37-rosettes and expressing CD7, CD5, CD2 antigens were characterized by the local activities of the above enzymes. The blasts did not express FcG-, FcM-, C3- or M-receptors. Cells like those at the Hodgkin disease and the "hand mirror" type cells were established on days 3-4. The number of lymphocytes with plasmatization was seen to increase by day 7.     

Differential expression of granulysin and perforin by NK cells in cancer patients and correlation of impaired granulysin expression with progression of cancer

Granulysin has been identified as an effector molecule co-localized with perforin in the cytotoxic granules of cytotoxic T lymphocytes and natural killer (NK) cells, and has been reported to kill intracellular pathogens in infected cells in the presence of perforin and to induce a cytotoxic effect against tumor cells. The aim of the present study was to elucidate whether intracellular expression of granulysin and perforin by NK cells might be associated with progression of cancer. Flow cytometric analysis demonstrated high levels of perforin and granulysin expression by CD3(-) CD16(+) cells in healthy controls. In contrast, cancer patients exhibited significantly decreased levels of granulysin expression ( P<0.005), despite having equally high levels of perforin expression in comparison with healthy controls. The tumor-free patients expressed granulysin at levels similar to healthy controls, while the progressive tumor-bearing patients expressed remarkably lower levels of granulysin compared to healthy controls ( P<0.0001). Similarly, patients with an advanced performance status had significantly fewer granulysin-positive NK cells than healthy controls. Meanwhile, a considerable number of the tumor-bearing patients showed a decrease in the number of circulating NK cells, and a correlation between impaired granulysin expression and reduced circulating NK cells was observed. These findings suggest that the tumor-bearing patients with impaired granulysin expression were in an immunosuppressive state. In conclusion, impaired expression of granulysin by NK cells correlates with progression of cancer, and determination of granulysin expression might prove informative for assessing the immunological condition of cancer patients.     

A single platform image cytometer for resource-poor settings to monitor disease progression in HIV infection.

BACKGROUND: For resource-poor countries, affordable methods are required for enumeration of CD4(+) T lymphocytes of HIV-positive patients. For infants, additional determination of CD4/CD8 ratio is needed. METHODS: We determine the CD4(+) and CD8(+) T lymphocytes as the CD3(+)CD4(+) and CD3(+)CD8(+) population of blood cells. Target cells are CD3-immunomagnetically separated from the whole blood, and CD4-Phycoerythrin and CD8-PerCP immunofluorescently labeled. A point-of-care single platform image cytometer was developed to enumerate the target CD3(+)CD4(+) and CD3(+)CD8(+) populations. It has light-emitting diodes illumination, is fully computer-controlled, operates from a 12 V battery, and was designed to be cheap and easy-to-handle. Target cells are imaged on a CCD camera and enumerated by an image analysis algorithm. The cytometer outputs the absolute number of CD4(+) and CD8(+) T lymphocytes/microl and CD4/CD8 ratio. RESULTS: The quality of the cell images obtained with the cytometer is sufficient for a reliable enumeration of target cells. The image cytometer achieves an accuracy of better than 10% in the range of 50-1700 cells/microl. Analysis of blood samples from HIV patients yields a good agreement with the TruCount method for CD4 and CD8 count and CD4/CD8 ratio. CONCLUSIONS: The image cytometer is affordable (component costs $3,000), compact (25 x 25 x 20 cm(3)), and uses disposable test materials, making it a good candidate to monitor progression of immunodeficiency disease in resource-poor settings.