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The immunological profile of B-cell disorders and proposal of a scoring system for the diagnosis of CLL.

We have investigated the role of immunophenotyping in distinguishing between leukemic B-cell lymphoproliferative disorders. Circulating cells from 666 cases were analyzed with a panel of markers by flow cytometry. The diseases included: chronic lymphocytic leukemia (CLL), 400; prolymphocytic leukemia, 22; hairy cell leukemia (HCL), 40; HCL variant, 15; splenic lymphoma with villous lymphocytes, 100; follicular lymphoma, 26; lymphoplasmacytic lymphoma, 25; mantle-cell lymphoma, 20; and large cell lymphoma, 18. On the basis of the most common marker profile in CLL, CD5+, CD23+, FMC7- and weak expression (+/-) of surface immunoglobulin (SmIg) and CD22, we devised a scoring system that gives for each of these five markers a value of 1 or 0 according to whether it is typical or atypical for CLL. Scores range from 5 (typical of CLL) to 0 (atypical for CLL). Application of the scoring system to all the cases showed that 87% of CLL scored 5 and 4 and only 0.4% scored 0 or 1, whereas 89% of other B-cell leukemias and 72% of lymphomas scored 0 or 1; only one case (0.3%) scored 4 and none scored 5 (p < 0.0001). There were no differences between CLL with high and low scores but higher scores were found in cases with more typical morphology (p < 0.0015). Considering each individual marker, there was no single one that distinguished CLL from other diseases, although the most reliable were SmIg intensity and FMC7. The proposed score will facilitate the diagnosis of B-lymphoproliferative disorders and improve their classification.

Antigens, CD↗

Immunological profile in congenital heart disease.

Fifty children with established congenital heart disease (CHD) were surveyed for the immune profile. Ventricular septal defect (VSD) was the commonest lesion (56%) followed by Tetralogy of Fallot (ToF; 16%), atrial septal defect (ASD; 8%), patent ductus arteriosus (PDA; 4%), transposition of great arteries (TGA; 4%), aortic stenosis (AS; 4%), and pulmonic stenosis (PS), tricuspid atresia (TA), single ventricle with pulmonic stenosis (SV with PS) and dextrocardia with ToF (2% each). Immunoglobulins (IgG, IgA and IgM) were estimated. IgG and IgA levels were significantly reduced in all children with congenital heart disease, whereas IgM levels were increased in cyanotic but unaffected in the acyanotic group. Complement C3 and C4 levels were reduced in all, more so in cyanotics. T-helper cells were decreased and T-suppressor cells were increased in all groups with congenital heart disease as compared to controls. B-cell percentage was increased in cyanotics but not affected in the acyanotics.

Antibody Formation↗

[Different immune profiles according to the immunological and clinical progression in vertically HIV-infected children].

BACKGROUND: Our goal was to evaluate immunologic profile differences of HIV-infected children on antiretroviral treatment (ART). PATIENTS AND METHODDS: We studied 23 HIV-vertically infected children: a) N-A1 group: 10 HIV-infected children in A1 category; b) N-B2 group: 6 HIV-infected children in B2 category, and c) N-C3 group: 7 HIV-infected children in C3 category. We also studied 13 healthy age-matched HIV-negative children as controls. Cell proliferation was evaluated by incorporation of [3H]-Thymidine. The cytokine production in culture was quantified using commercially available specific ELISA assays. T-cell subsets were determined by flow cytometry. RESULTS: Stimulation indexes of PHA, PWM, and anti-CD3+ anti-CD28 in N-A1 group were higher than in N-C3 group. In unstimulated PBMC, TNF-alpha production of HIV-infected children was higher than the control group (p < 0.05). However, in stimulated PBMC, TNF-alpha production in N-B2 and N-C3 groups was lower than the control group (p < 0.05). In HIV-infected children, CD8+ CD45RA+ CD62L+ T-cells were significantly lower (p < 0.01) and CD8+ CD45RO+ T-cells were higher (p < 0.05) than the control group. Moreover, in NA-1 group, CD4+ CD45RA+ CD62L+ T-cells were higher, and CD4+ CD45RO+ and CD8+ CD45RO+ T-cells were lower, than in N-B2 and N-C3 groups (p < 0.05). On the other hand, CD45RO+, CD45RO+ CD38+, HLA-DR+, CD38+ HLA-DR+ and CD38+ CD4+ and CD8+ T-cells were higher in N-C3 group than the N-A1 and control groups, except for CD4+ CD38+ T-cells. Activated CD8+ T-cells in N-A1 group were higher than in control group (p < 0.01). CONCLUSION: Our data demonstrate that in spite of ART, there still remain important differences in the immunologic status of HIV-infected children depending on the HIV-infection stage.

CD4-CD8 Ratio↗

[Evaluation of immunological studies in patients with rheumatoid arthritis].

Immunological studies are widely applicable at a clinic in rheumatoid patients, despite the fact that their clinical value is still a disputable subject. Many authors are of opinion that the determination of individual immunological tests does not provide the estimation of immunological state in patients, and should be evaluated critically. The aim of the actual paper was to evaluate the immunological state of patients with rheumatoid arthritis by means of immunological profile index, which consists in performing concurrently many immunological tests involving both humoral immunology and cellular one. The results were referred to the duration of the disease, pathological process activity, the presence of rheumatoid factor, the advancement of osseous changes. The studies were carried out in patients, aged 16-69 years, 63 of them had RA and 9 were affected by ankylosing spondylitis. The control group comprised 16 normal subjects aged 28-61 years. It has been disclosed that the values of the immunological profile index in patients were statistically significantly lower than in the control group. The lowest values of immunological profile index were recorded in patients with extra-articular symptoms with recurrent infections of urinary and respiratory tracts, as well as in patients with active form of RA, and the presence of rheumatois factor. The patients with ankylosing spondylitis were found to reveal immunological disorders, but they were expressed less markedly than in RA patients. With the help of immunological profile it was possible to show that multifunctional immunological disorders appeared in patients with rheumatoid arthritis. The most sensitive immunological tests were: levels of immunoglobulins in blood serum, and the count of lymphocytes T and B. The immunological profile study in patients with rheumatoid inflammation of joints is an objective method of evaluating their immunological state.

Adolescent↗

Understanding clinical immunological testing in alleged chemically induced environmental illnesses.

Some believe that an abnormal immunoregulatory response based on environmental damage to T cells is fundamental to the production of symptoms in patients with alleged "multiple chemical sensitivity" and/or "environmental illness." According to this theory stimulation of T cells or T cell phenotypic subsets by environmental chemicals results in release of cytokines that can effect appropriate target cells of multiple organ systems, resulting in a wide range of symptoms. This concept is reinforced by frequent media reporting of pollution incidents and environmental disasters plus continued isolated reports of immunologic abnormalities in patients with various forms of alleged environmental illness, multiple chemical sensitivities, or other related syndromes. These include reports of slight perturbations in quantity and function of immunoglobulins, complement and its components, B cells, natural killer cells, T cells, phenotypic T cell subsets, and helper suppressor T cell ratios. There are also reports of increased or decreased interleukin levels including IL-1 and IL-2 or their receptors (IL-2R) in these patients. Such assays are not infrequently performed even though there is no evidence for their diagnostic efficacy in these alleged conditions. It is reasonable, however, to anticipate that with the wide development of assays for many of the interleukins and their receptors, these assays may become important in the future diagnosis of many autoimmune, allergic, neoplastic, and infectious diseases. At this time, however, the induction of environmental illness or multiple chemical sensitivity by exposure to trace levels of environmental "immunotoxins" is unproven and remains a matter of speculation. The reproducibility of immunologic test abnormalities reported under these conditions has not been documented, and the data have often not been analyzed statistically. Appropriate controls also have not usually been employed, nor have control values been provided in many cases. Without consideration of these factors, a patient might be erroneously diagnosed as having some form of "immune dysregulation," "environmental immune dysfunction," or "immunotoxic" syndrome on the basis of only a single panel of cellular immunologic profiles or related immunologic tests illustrating slight deviations from the norm and in the absence of overt disease on physical examination. Consideration must also be given to an understanding of biologic variability and diurnal variations in lymphoid cell numbers in interpreting cellular immunologic profiles. For example, the necessity for age and sex-matched controls, test reproducibility, quantitative versus functional assays, and the significance of major versus minor deviations from the norm must be appreciated. In addition, many other conditions can effect immunologic tests, such as medications, psychologic factors, cigarette smoking, and the presence of concurrent disease, including minor viral infections. All of these variables should be appreciated in test interpretation. Certain clinical indications for analysis of cellular components of the immune system, using flow cytometry, have been provided as guidelines although they are by no means accepted by all groups due to their current incomplete evaluation by the clinical immunology community. These suggested indications are discussed. In this article, attempts are made to outline the various quantitative and functional tests used to assess the immune system, with emphasis on "biomarker" tests to detect possible immune system "damage." Dangers involved in attempting to make clinical evaluations based on results of isolated in vitro assessment of quantity or function of immune system cellular and humoral components without considering the results of a good medical history and physical examination, the many pitfalls involved in the tests, and the many confounding variables that affect the tests are emphasized, as well as the need for proper controls...

Autoantibodies↗

Gene expression profile in immunologically injured liver cell of mice.

To study the gene expression profiles between immunologically injured liver cell and normal liver cell of mice and to screen on a large scale the differentially expressed genes associated with the formation of liver injury, the experimental mice were randomly divided into the normal group for controlling and the immunologically liver-injured group induced by BCG and LPS. The liver mRNA of the two groups were extracted respectively and reversely-transcribed to cDNA with the incorporation of different fluorescence (Cy3, Cy5) labeled dUTP as the hybridization probes. The mixed probes were hybridized to the cDNA microarray chips. The fluorescent signal results were acquired by scanner ScanArray 4000 and analyzed with software GenePix Pro 3.0. Among the 14112 target genes, 293 genes were found to be significantly differentially expressed, in which 188 genes were up-regulated and 105 genes were down-regulated. Based on the analysis of biological functions of those differentially expressed genes, it was indicated that the occurrence and development of mouse liver damage induced by BCG and LPS were highly correlated with the processes of immune reactions, cell synthesis, metabolism, apoptosis and transportation in liver cell, which might be quite important for elucidating the regulatory network of gene expression associated with the liver damage, also important for finally discovering the pathogenic mechanisms of immunological liver damage.

Alanine Transaminase↗

[Preliminary data on the study of various immunologic parameters in hemophilic children].

Treatment of coagulation deficiencies with high dosages of the missing factor is a source of continuous diffusion of homologous proteins that could modify the normal immunological profile. We have performed immunologic studies on 24 children with ages ranging from 1 - 16 years with severe classical hemophilia and 1 child with von Willebrand's and on a control of 19 age-matched healthy children. Our preliminary data show no statistically significant alterations in the basic immunological profile between normal children and those with hemophilia. Nor were there any differences among the hemophilic patients on the various transfusional regimes. Considerations are made on the reasons for the discrepancies between our results and those on adults presented in the literature.

Adolescent↗

TH2 profile in asymptomatic Taenia solium human neurocysticercosis.

Neurocysticercosis (NC), a parasitic disease caused by Taenia solium, may be either asymptomatic or have mild to severe symptoms due to several factors. In this study, the immunological factors that underlie NC pleomorphism were studied. Ten of the 132 inhabitants of a rural community in Mexico (Tepez) had a computerized tomography (CT) scan compatible with calcified NC, and all were asymptomatic. Their immunological profiles were compared with those of 122 CT scan negative (non-NC) subjects from the same village. NC was associated with a TH2 response (IgG4, IL-4, IL-5, IL-13). Subjects from Tepez had higher levels of specific antibodies (IgG1, IgG2, IgG4, IgE) and specific cell proliferation than subjects from an area with low exposure (Ensenada). This suggests that non-NC subjects from Tepez had been exposed to T. solium and resisted infection in the brain. Distinct immunological profiles in equally exposed individuals differing in outcome of infection support the hypothesis of host-related factors in resistance to and pathogenesis of NC. This is the first study reporting the immunological profile associated with the asymptomatic form of NC.

Adolescent↗

Single cell studies on the immunological marker profile of plasmacytoid T-zone cells.

Plasmacytoid T cells (PTC) are known to home to thymic (T) zones in human lymph nodes and are characterized by their abundant, concentrically layered, rough endoplasmic reticulum. These cells have been found in reactive and neoplastic conditions. Three cases of PTC lymphomas have so far been reported. All of them were complicated by a myelomonocytic leukemia leading to the assumption of a functional relationship between PTC and the myeloid system. The immunologic phenotype of PTC, as revealed on frozen tumor tissue sections, comprised the expression of CD5 (T1), CD4 (T4), and HLA-DR, but not CD8 (T8) and CD2 (T11) and suggested an affiliation to the T cell system. Extending our previous report on one of these cases we here present the first study on the immunological marker profile of suspended PTC. The employment of unfractionated or PTC-enriched tumor cell suspensions rendered possible the application of a panel of monoclonal antibodies (moAbs) on both fixed and unfixed cells and enabled us to allocate various markers either to the intracytoplasmic or surface domain of this cell type. Our results suggest that PTC from our case rest in the G0/G1 phase of the cell cycle. They express the transferrin receptor, but not the Il-2 receptor (CD25) or the nuclear antigen Ki-67. No T cell antigen was demonstrated on the surface of unfixed suspended PTC. Under these conditions only HLA-DR and a predominantly monocytic antigen (CD36/moAb 5F1) were identified. Fixed cells, however, showed a weak cytoplasmic reactivity for CD5 and two myelomonocytic antigens (CD15/moAb 1G10 and CD14/moAb My4). Our findings do not sustain positive evidence for a T cell nature of PTC. Whether their phenotypical pattern indicates terminal differentiation with concomitant loss of T cell antigens or points to a cytogenetic relationship of PTC to the myeloid system, remains speculative. Until the cytogenesis of PTC is clarified we propose the noncommitted term "plasmacytoid T-zone cells" for this elusive cell type.

Antigens, Differentiation, T-Lymphocyte↗

Focal stiff-person syndrome.

Stiff-person syndrome (SPS) is a disorder of motor function characterized by rigidity of axial musculature and fluctuating painful spasms, which are often induced by startle or emotional stimuli. Neurophysiological studies have demonstrated the presence of continuous motor unit activity in muscle at rest, with abnormally enhanced extereoceptive reflexes. Although criteria for the diagnosis of SPS were proposed, several variants of this syndrome have been described before. In this communication, we report the case of a patient with a focal form of SPS. A 39-year-old woman developed progressive instability in her gait, spasms and stiffness restricted to both legs. The electromyographic examination showed continuous motor unit activity of the affected muscles at rest. Moreover, high anti-GAD antibodies titers were found in CSF and serum. Clinical symptoms, electrophysiological and immunological profiles suggest a focal form of SPS. Clinical and immunological findings indicate that SPS is a heterogeneous disease, suggesting the need to redefine its diagnostic criteria. Definition of the range of clinical expression and immunological profiles could be important for the clinical management of these patients.

Adult↗

Hormonal profiles and immunological studies of male lupus in Taiwan.

The aims of this study were to describe hormonal profiles, cytokine production and Fc-gamma receptor (Fcgamma-R) distribution in male lupus patients in Taiwan, and to look for any differences between our patients and normal individuals. Sixteen newly diagnosed and untreated male lupus patients were studied. Hormonal profiles were determined by radioimmunoassay. Interleukin-1 (IL-1) and IL-1 receptor antagonist (IL-1ra) production from both monocytes and neutrophils was determined by ELISA and murine thymocyte proliferation assay. The FcgammaR distribution on both monocytes and neutrophils was detected by flow cytometer. There were no significant differences in FSH, LH, testosterone, oestradiol, and beta-HCG blood levels in male lupus patients compared with normal individuals; however, the prolactin level in lupus patients was significantly higher than in normal individuals. Furthermore, there was no difference in IL-1 and IL-1ra production from both monocytes and neutrophils among male and female lupus patients, and normal individuals. Male lupus patients have a significantly lower FcgammaRII distribution on both monocytes and neutrophils when compared with female lupus patients and normal individuals. It was concluded that the high prolactin level and low FcgammaR distribution may play a role in the pathogenesis and prognosis of male lupus.

Adult↗

Validation of a Pseudomonas aeruginosa porcine model of septic shock.

OBJECTIVES: To develop a standardized bacteraemic porcine model of septic shock with cardiovascular and immunological profiles similar to those observed in human clinical states. METHODS: Sepsis was induced by an intravenous challenge of 18 anaesthetized pigs with live Pseudomonas aeruginosa. The pulmonary arterial pressure was monitored and the bacterial infusion was stopped when the systolic pulmonary arterial pressure reached 45 mmHg. Septic shock was treated with fluid resuscitation and epinephrine infusion. The haemodynamic parameters and the rate of different inflammatory cytokines were recorded during 6 h of observation. RESULTS: The mean+/-SD cardiac output increased from 2.4+/-1.2 to 5.7+/-2.1 L/min while the mean+/-SD systemic vascular resistance index decreased from 1957+/-744 to 709+/-221 dyn/s/cm5/m2. The pharmacokinetic profile of the inflammatory cytokines was similar to the one observed in human studies. CONCLUSIONS: The control of the systolic pulmonary arterial pressure during a P. aeruginosa infusion leads to a hyperdynamic, reproducible cardiovascular profile similar to the one observed in human septic shock. Since the immunological profile of the inflammatory cytokines is also similar to the human one, this standardized porcine model appears to be appropriate for experimental research concerning sepsis.

Animals↗

Profile of systemic sclerosis in a tertiary care center in North India.

AIM: To study the clinical and immunological profile in patients of systemic sclerosis from North India and compare it with other ethnic groups. METHODS: Patients presenting to us between the years 2001 and 2004 and fulfilling the American Rheumatism Association (ARA) criteria for systemic sclerosis were included. There were 84 females and 16 males with the mean age of 32.5 +/-11.62 years and a mean duration of 6.49 +/- 4.34 years. All patients were admitted to the dermatology ward for detailed history and examination including Rodnan score. Investigations including hemogram, hepatic and renal functions, serum electrolytes, urine for albumin, sugar, microscopy and 24h urinary protein estimation, antinuclear antibody, chest X-ray, barium swallow, pulmonary function test, electrocardiogram and skin biopsy were done. RESULTS: The most common presenting symptoms were skin binding-down (98.5%), Raynaud's phenomenon 92.9%, pigmentary changes 91%, contracture of fingers 64.6%, fingertip ulcer 58.6%, restriction of mouth opening 55.5%, dyspnea 51.1%, joint complaints 36.7% and dysphagia in 35.2%. The mean Rodnan score was 25.81 +/- 10.04 and the mean mouth opening was 24.6 +/- 19.01 mm. The laboratory abnormalities included raised ESR in 87.8%, ANA positive in 89.1%, proteinuria in 6.0%, abnormal chest X-ray in 65.3%, abnormal barium swallow in 70.2% and reduced pulmonary function test in 85.8%. CONCLUSION: The clinical and immunological profile of systemic sclerosis in North India is similar to that of other ethnic groups except that pigmentary changes are commoner and renal involvement is relatively uncommon.

Adolescent↗

Serologic profiles as immunologic markers for different clinical presentations of lupus erythematosus.

The clinical and laboratory features of 55 patients with lupus erythematosus (LE), grouped on the basis of six nuclear immunofluorescent pattern results commonly encountered in this disease were examined. Serologic profiles of antinuclear antibodies (ANA), anti-DNA and anti-ENA results can serve as immunologic markers in LE for a benign subset and two other groups with a different incidence of certain clinical characteristics. The large speckle-like thready pattern without antibodies to DNA or ENA is an immunologic marker for a benign LE subset, with generalized skin lesions with or without joint involvement only. Significant levels of the anti-DNA antibodies with the shrunken peripheral, peripheral, or leukocyte-specific ANA with a particulate pattern are markers for severe systemic involvement. The thready pattern with antibodies to ENA (Sm antigen) and leukocyte-specific ANA without a particulate pattern, with or without antibodies to DNA or ENA, indicate less severe systemic disease.

Adolescent↗

Assessment of hematological and immunological function during long-term follow-up after peripheral blood stem cell transplantation.

BACKGROUND AND OBJECTIVE: Long-term hemopoietic and immunological profile after autologous peripheral blood progenitor cells transplantation (PBPCT), in patients affected by hematological malignancies is largely unknown. The aim of this work was to detect the impact of high dose chemotherapy and PBPCT on hemopoietic and immunological function compared to conventional chemotherapy. DESIGN AND METHODS: Patients had to fulfill the following criteria: continuous complete remission after PBPCT, follow-up longer than 12 months, no chemo or radiotherapy or biological response modifiers after PBPCT. Twenty-five patients were considered eligible for this analysis. Stable and complete hemopoietic reconstitution (Hb > 12 g/dL, WB > 4.0 x 10(9)/L, ANC > 1.5 x 10(9)/L and Pits count > 150 x 10(9)/L), morphological examination of peripheral blood and bone marrow, cytogenetic analysis and immunological profile were evaluated at 12 months and yearly thereafter. RESULTS: Immunological reconstitution showed a persistent reduction of CD4/CD8 ratio up to five years after PBPCT. This reduction was related to a persistent increase of CD8+ lymphocytes and a constant reduction of CD4+ lymphocytes. INTERPRETATION AND CONCLUSIONS: Defects observed in PBPCT patients are induced by the procedure itself, by the conditioning regimen or both. The different behavior in the immune reconstitution of CD8+ subset after PBPCT may be favored by an extrathymic origin of these cells while CD4+ subset recovery which is thymus-dependent is impaired after PBPCT in adult population. Long-term hemopoietic reconstitution after PBPCT is rapidly obtained and is stable over the years, long-term immunological function seems to be abnormal in these patients and these abnormalities are long-lasting.

Adult↗