Association of HLA antigens with certain diseases provide insights into genetically determined susceptibility to disease. Although nephrotic syndrome is one of the commonest diseases, it is poorly understood. A group of 57 patients suffering from a minimal lesion nephrotic syndrome (33 patients) and mesangioproliferative glomerulonephritis (24 patients) was studied for immunologic markers. The incidence of HLA-A w 24 is significantly greater in the minimal lesion nephrotic syndrome patients than in controls (18.7% in patients, 0% in controls, p < 0.01). This report fails to show a high incidence of specific HLA antigen in mesangioproliferative glomerulonephritis patients. We believe that the high incidence of HLA-Aw 24 in minimal lesion nephrotic syndrome is indicative of a congenital predisposition to nephrotic syndrome.
Glomerulonephritis/immunology ; HLA Antigens/analysis* ; Human ; Nephrosis, Lipoid/immunology*

A 43-year-old man with generalized lichen planus demonstrated serum antibodies against autologous lesional skin. Indirect immunofluorescence using serum and papular lesional skin revealed a lichen planus specific antigen found only in the granular layer. The specific tissue antigen was not detected in normal skin from this patient, in normal skin from patients with skin disorders other than lichen planus or in skin from normal control persons. When titers of the serum antibodies against lichen planus antigen were examined before and after a successful therapy a positive correlation of the titer could be found in this patient.
Adult ; Antibodies/*immunology ; Antigens/*analysis ; Humans ; Lichen Planus/drug therapy/*immunology ; Male ; Skin/*immunology

To investigate the genetic factors in Koreans with leprosy, 157 unrelated leprosy patients have been typed for HLA antigens, and compared with 162 healthy controls. The patient group consisted of 124 with lepromatous leprosy and 33 with tuberculoid leprosy. HLA-A11 was found to be increased in lepromatous leprosy (p=0.0005). HLA-Aw33 was found to be increased in both lepromatous leprosy (p = 0.0002) and tuberculoid leprosy (p = 0.005). HLA-Cw5 was found to be decreased in lepromatous leprosy (p = 0.009). Frequencied of HLA-B antigens did not differ significantly between the leprosy patients and the healthy controls.
HLA Antigens/analysis* ; Human ; Korea ; Leprosy/genetics ; Leprosy/immunology* ; Phenotype

OBJECTIVETo research the maturation regulation of dendritic cells (DCs) pulsed with hepatocellular carcinoma (HCC) cell soluble antigens.

METHODSBCG HSP 70 was purified by SDS-PAGE electrophoresis and its biological activity was determined with ELISA. Phenotypes of DCs pulsed with antigens or with both antigens and BCG HSP 70 were analysed with flow cytometry. MTT assay was used to estimate the proliferation of self lymphocytes and the mixed lymphocyte reaction (MLR) of BCG HSP 70 primed DCs.

RESULTSThe characteristics of DCs had changed after loaded with soluble antigens of HCC. There were about 10% DCs which had lost their specific markers. The expression levels of CD54, CD83, CD86 molecules and the stimulatory ability in allogeneic MLR decreased. However, after being activated by BCG HSP 70, the DCs pulsed with antigens could keep their special markers and the expression levels of CD54, CD83, CD86 molecules increased too. The stimulatory abilities in allogeneic MLR and proliferation of self lymphocytes also improved.

CONCLUSIONThis study shows that BCG HSP 70 can induce DCs pulsed with antigens maturation and improve their antigen-presenting ability, which may be a useful maturation inducer for dendritic cells.

Antigen Presentation ; Antigens, CD ; analysis ; Antigens, Neoplasm ; immunology ; B7-2 Antigen ; Carcinoma, Hepatocellular ; immunology ; Dendritic Cells ; cytology ; immunology ; HSP70 Heat-Shock Proteins ; immunology ; Humans ; Immunoglobulins ; analysis ; Intercellular Adhesion Molecule-1 ; analysis ; Liver Neoplasms ; immunology ; Membrane Glycoproteins ; analysis ; Mycobacterium bovis ; immunology

The aim was to investigate the cytolytic activity of cytotoxic T lymphocytes (CTL) induced by dendritic cells (DC) derived from human cord blood in vitro. Human cord blood mononuclear cells (CBMNC) were cultured in vitro with addition of various cytokines. DC was confirmed by morphology, immune phenotype and capacity of stimulating MLR (mixed lymphocyte reaction). CTL were generated through the co-culture of autologous T lymphocytes and DC. (51)Cr-release assays were performed for the measurement of cytotoxicity of CTL. The results showed that distribution of the subgroups of T lymphocytes in CBMNC was similar to that in adult peripheral blood. The percentage of CD1a-expressing cells in CBMNC was very low, merely (0.41 +/- 0.09)%. During culture, DC became larger and more irregular in shape. Spiny dendrites or multiple cell processes in morphology emerged on the surface of DC. Among the cell populations at 15 days of culture, there were (28.4 +/- 3.55)% of CD1a-expressing cells, (63.67 +/- 23.33)% of CD86-positive, (8.7 +/- 1.49)% of CD83-positive and (32.5 +/- 1.53)% of HLA-DR-positive cells. DC derived from CBMNC is capable of stimulating the proliferation of allogeneic lymphocytes in MLR. CTL derived from autologous T lymphocytes induced by dendritic cells pulsed with lysates of HL-60 cells, possessed specific cytolytic effects against HL-60 cells. In conclusions, relatively high percentage of CD1a-expressing cells can be generated in culture system of this study. DC derived from cord blood is able to induce the production of anti-leukemic CTL in vitro.
Antigens, CD1 ; analysis ; Cytotoxicity, Immunologic ; Dendritic Cells ; immunology ; Fetal Blood ; immunology ; Humans ; Immunophenotyping ; Leukemia ; immunology ; T-Lymphocytes, Cytotoxic ; immunology

ZCH-2B8a (IgG2a) is a novel monoclonal antibody (McAb) generated in laboratory of Children Hospital of Medical College, Zhejiang University recently using human myeloblastic leukemia cell line KG1a as immunogen. This antibody has been submitted to the 8th International Workshop and Conference on Human Leukocyte Differentiation Antigens (HLDA8) and the results showed that the antibody recognized an unknown molecule on the surface of some blood cells. The aim of this study was to investigate the reactivity of this antibody on normal blood cells and malignant cell lines and to explore its possible application in clinical practice. The multi-parameter flow cytometry was used to analyze the expression pattern of 2B8a antigen in triplicate on normal blood components including T cells, B cells, natural killers (NK), neutrophils, monocytes, dendritic cells (DC), red blood cells (RBC), platelets (Plt), hematopoietic stem/progenitor cells derived from either bone marrow or G-CSF mobilized peripheral blood CD34(+) cells and malignant cell lines including 14 hematopoietic, 5 neuroblastoma, 1 colon cancer and 1 amniotic epithelium cell lines. The amount of positive cells > or = 20% was considered as positivity. The results showed that 2B8a antibody reacted to 3/3 specimens of blood B cells with a positive rate of 26.29% and 2/3 specimens of monocytes with an average positive rate of 59.84%. 2B8a was weakly reactive to neutrophils (23.72%) and negative for T cells, NK, DC, RBC and Plt. The antibody reacted to all 3 marrow CD34(+) cells with an average positive rate of 39.33% while it was negative for G-CSF-mobilized CD34(+) peripheral blood stem/progenitor cells (PBSC, 1.25%). Cell line analysis showed that the antibody notably reacted to three out of 4 cell lines (Raji, SMS-SB, Nalm-6 and Nall-1) with the positive rates of 98.78%, 98.61%, 94.93% respectively and weakly to one of them with 5.68% in B lineage cell lines and monoblastic cell line (U937, 67.78%) while it was only weakly positive or negative for other myeloid leukemia cell lines including Meg01 (33.40%), HL-60 (29.70%), K562 (28.19%), KG1a (16.23%) and HEL92.1.7 (8.02%). Among 4 T lineage leukemia, 5 neuroblastoma and 1 colon cancer cell lines tested, only Molt-3 was found weakly positive (31.40%) for 2B8a, while the remaining 3 T cell lines (Molt4, JM and CCRF-CEM), 5 neuroblastoma cell lines (LA-N1, KCNR, BE, SK-N-SH, SK-N-AS) and the colon cancer cell line (HR8348) tested were negative. An amniotic epithelium cell line (FL) was showed positive for the antibody (45.03%). It is concluded that 2B8a antibody primarily reacts to B lineage and monocytic lineage cells which may bear the diagnostic and therapeutic applications among different types of hematopoietic malignancies.
Antibodies, Monoclonal ; biosynthesis ; immunology ; Antibody Specificity ; Antigen-Antibody Reactions ; Antigens, Neoplasm ; analysis ; immunology ; Antigens, Surface ; immunology ; B-Lymphocytes ; cytology ; immunology ; HLA Antigens ; immunology ; Hematopoietic Stem Cells ; cytology ; immunology ; Hematopoietic System ; cytology ; Humans ; Leukemia, Myeloid, Acute ; immunology ; pathology ; Tumor Cells, Cultured

OBJECTIVETo explore the clinicopathologic features, immunophenotype, differential diagnosis and gene mutation status of the Erdheim-Chester disease (ECD).

METHODSClinical and pathologic findings of 3 ECD cases were examined by gross, microscopic, immunohistochemical methods and BRAF V600E mutation. Related literatures were reviewed.

RESULTSTwo male patients and one female patient presented clinically with multiple skin nodules, bone pain and bony lesions by imaging study. Microscopically, the lesions were composed of spindle-shaped fibroblasts, foamy histiocytes and scattered Touton-type giant cells embedded in reactive fibrous tissue. Lymphocytes, plasma cells, and multinucleated giant cells were also found. Immunohistochemically, all histiocytes were positive for CD68, none of which expressed CD1a, although 2 cases focally expressed weak S-100 stain. In 2 cases,BRAF V600E mutation was detected.

CONCLUSIONSECD is a rare disease of xanthogranulomatous histiocytosis.Its diagnosis relies on pathological and immunohistochemical findings, but correlation with clinical information, especially radiographic findings should be performed.No effective treatment of the disease is currently available.

Antigens, CD ; analysis ; Antigens, CD1 ; analysis ; Antigens, Differentiation, Myelomonocytic ; analysis ; Diagnosis, Differential ; Erdheim-Chester Disease ; genetics ; immunology ; pathology ; Female ; Humans ; Male ; Mutation ; S100 Proteins ; analysis ; Treatment Outcome

PURPOSE: Tuberculosis (TB) is a major infectious disease and is responsible for two million deaths annually. For the identification and quantitation of Mycobacterium tuberculosis (M. tuberculosis), a causative agent of TB, a sandwich enzyme-linked immunosorbent assay (ELISA) against the MPT64 protein of M. tuberculosis, an antigen marker of the M. tuberculosis complex, was developed. MATERIALS AND METHODS: The MPT64 protein was expressed, and anti-MPT64 monoclonal antibodies were prepared. A sandwich ELISA was established using recombinant MPT64 protein and anti-MPT64 monoclonal antibodies. The sandwich MPT64 ELISA was evaluated using reference and clinical mycobacterial strains. RESULTS: The sandwich MPT64 ELISA detected MPT64 protein from 2.1 ng/mL to 250 ng/mL (equivalent to 1.7x10(4) CFU/mL and 2.0x10(6) CFU/mL). All 389 clinical M. tuberculosis isolates tested positive in the sandwich MPT64 ELISA (sensitivity, 100%), and the assay showed no cross reactivity to any tested nontuberculous mycobacterial strain (specificity, 100%). CONCLUSION: The sandwich MPT64 ELISA is a highly sensitive and quantitative test for MPT64 protein, which can identify M. tuberculosis.
Antigens, Bacterial/*analysis/immunology ; Enzyme-Linked Immunosorbent Assay/*methods ; Mycobacterium tuberculosis/*immunology

OBJECTIVETo investigate the possibility of using melanoma antigen-1 (MAGE-1) peptide as a tumor vaccine to treat hepatocellular carcinoma (HCC).

METHODSThe expressions of MAGE-1 in 8 HCC cell lines and in liver cancer tissue from a patient were detected using RT-PCR. The type of human leucocyte antigen I(HLA I) of both 8 HCC cell lines and peripheral blood mononuclear cells of the patient was detected using a microcytotoxicity method to screen out target cell lines for the cytotoxicity assay. Peripheral blood mononuclear cells from the HCC patient pulsed with an MAGE-1 peptide (NYKCRFPEI) were used as antigen presenting cells. Autogenous peripheral blood mononuclear cells were stimulated with antigen presenting cells every 7 days for 4 times to elicit cytotoxic T lymphocytes. The phenotype of effector cells was analyzed using flow cytometry. The cytotoxicity of effector cells was detected with a lactate dehydrogenase releasing assay.

RESULTSThe expressions of both MAGE-1 and HLA-A24 were detected in BEL7405 cell line which were used as the positive target cell line in the cytotoxicity assay. The expression of MAGE-1 alone was detected in HLE, BEL7402, BEL7404, QGY7703 and SMMC7721 cell lines, and the expression of neither MAGE-1 nor HLA-A24 was shown in QGY 7701 and HpG2 cell lines. The last 7 cell lines could be used as negative target cell lines in the cytotoxicity assay. Peripheral blood mononuclear cells expanded 32 folds during 28-day culture. The ratio of CD3(+) T cells increased by 16% (from 54% to 70%), and the ratio of CD8(+) T cells increased by 20% (from 36% to 56%) during 28-day culture. When the ratio of effector cells to target cells was 10:1, effector cells exhibited 62.5% cytotoxicity against autogenous lymphoblasts pulsed with the peptide (NYKCRFPEI) of MAGE-1 antigen, 40.25% cytotoxicity against BEL7405 cells, compared with 17.88% cytolysis observed against autogenous lymphoblasts, 19.55% against HLE cells, and 1.6% against QGY7701 cells. When the ratio of effector cells to target cells was 3.3:1, the cytotoxicity of effector cells against the peptide pulsed autogenous lymphoblasts was 53.6%, which was much higher against autogenous lymphoblasts, HLE cells and QGY7701 cells at 15.6%, 13% and 1%, respectively.

CONCLUSIONThe results demonstrate that cytotoxic T lymphocytes with the ability to specifically lyse target cells expressing both MAGE-1 and HLA-A24 could be successfully induced by the MAGE-1 peptide NYKCRFPEI in vitro. This indicates that a good result might be anticipated if this peptide is used as a tumor vaccine to treat HLA-A24 HCC patients.

Adult ; Antigens, Neoplasm ; Cancer Vaccines ; immunology ; Carcinoma, Hepatocellular ; immunology ; HLA-A Antigens ; analysis ; HLA-A24 Antigen ; Humans ; Liver Neoplasms ; immunology ; Male ; Melanoma-Specific Antigens ; Neoplasm Proteins ; genetics ; immunology ; RNA, Messenger ; analysis ; T-Lymphocytes, Cytotoxic ; immunology ; Tumor Cells, Cultured

OBJECTIVETo analyse the expression of glycosylphosphatidylinositol (GPI)-anchored protein and functional transition of lymphocyte subsets in paroxysmal nocturnal hemoglobinuria (PNH) patients.

METHODSFlow cytometer and multi-color McAbs were used to detect CD(59) expression on lymphocytes and their subsets. Stimulation test was used to examine the ability of affected T cells in converting from CD(45RA) phenotype to CD(45RO) phenotype.

RESULTSIn 14 PNH samples, CD(59) expression on affected lymphocytes was at a lower percentage than that on granulocytes and erythrocytes. B lymphocytes were widely affected than NK or T cells. CD(59)(-) deficient B lymphocytes existed in all PNH patients. More CD(8)(+) T cells were affected than CD(4)(+) T cells. Expression of HLA-DR and CD(45RO) on affected T cells were significantly lower than that on normal T cells (P < 0.01), whereas the expression of CD(45RA) was much higher on the former (P < 0.01). After stimulation with phytohemagglutinin (PHA), the expression of CD(45RA)(+) on affected T cells decreased sharply (the median value was from 82.7% to 26.6%), while the expression of CD(45RO)(+) increased significantly (the median value was from 19.6% to 64.3%).

CONCLUSIONSThe deficient CD(59) phenotype does involve in the different types of lymphocytes in PNH patients. Therefore affected B cells could be a good marker for diagnosis of PNH. The affected T cells were mainly comprised naive cells (CD(45RA)(+)HLA-DR(-)), which could acquire the memory phenotype (CD(45RO)(+)HLA-DR(+)) after stimulated by antigen.

Adolescent ; Adult ; CD59 Antigens ; analysis ; Female ; Hemoglobinuria, Paroxysmal ; blood ; immunology ; Humans ; Leukocyte Common Antigens ; analysis ; Lymphocyte Subsets ; cytology ; immunology ; Lymphocytes ; cytology ; immunology ; Male ; Middle Aged