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

This study was aimed to investigate the value of CD58 in evaluation of early therapeutic effect on childhood B-ALL. The expression features of CD58 in 135 cases of childhood B-ALL were analyzed by four-color flow cytometry; MRD detection protocol for B-ALL using CD58/CD10/CD34/CD19 combination was established; the correlation between the expression features of CD58 and MRD detection was analyzed for the early therapeutic response in childhood B-ALL. The results showed that the mean value of CD58 MFI in 135 cases of B-ALL was 113.08 +/- 63.33, which was significantly higher than that in 15 cases of normal bone marrow controls (14.68 +/- 5.26, P < 0.01). In addition, CD58 was over expressed in 51.9% (70/135) of B-ALL patients, indicating that CD58 could be an effective marker in MRD detection. The CD58/CD10/CD34/CD19 was the second most effective combination next to TdT/CD10/CD34/CD19 in B-ALL MRD detection with flow cytometry. Meanwhile, the positive rate of MRD detection by flow cytometry was significantly lower in CD58 over expression group (P < 0.05). It is concluded that CD58 may be used as an indicator for detection of MRD in B-ALL patients, which would enrich the combination of MRD detection. The CD58 over expression may be considered as a marker of a favorable prognosis in childhood B-ALL.
Biomarkers, Tumor ; analysis ; Burkitt Lymphoma ; immunology ; pathology ; CD58 Antigens ; analysis ; Child ; Humans ; Neoplasm, Residual ; Prognosis

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

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 investigate the relationship between six-transmembrane epithelial antigen of the prostate (STEAP) expression and the histologic grading of prostatic carcinoma (PCa).

METHODSDifferent prostatic tissues and non-prostatic tumors, 131 cases of PCa, 164 cases of benign prostate hyperplasia (BPH), and 56 cases of non-prostatic malignancies, were analyzed for the expression of STEAP by using STEAP monoclonal antibody and SP immunohistochemical staining. The positive area unit (PU) was introduced to describe the intensity of STEAP expression.

RESULTSThe prostatic tissues of all but 3 cases of PCa and 5 cases of BPH were stained positively, while all of the non-prostatic tumors were stained negatively. There was a significant negative correlation between the STEAP expression and the histologic grading of PCa.

CONCLUSIONSTEAP can be a prognostic marker of PCa and a potential therapeutic target in PCa.

Antigens, Neoplasm ; analysis ; genetics ; Humans ; Immunohistochemistry ; Male ; Oxidoreductases ; Prostatic Neoplasms ; diagnosis ; immunology ; pathology

OBJECTIVETo verify the obtained immunogenic membrane antigens candidate of pancreatic cancer in the performed research.

METHODSPancreatic cancer cell line SW1990 membrane protein underwent immunoblot with serum IgG purified from clinically collected sera of 66 pancreatic cancer patients. Number 3 and number 8 positive dots of immunoblot were identified by MALDI-TOF mass spectrometry and peptide mass fingerprinting matching. The candidate membrane antigens were further validated in cell lines by RT-PCR, real-time PCR and Western blot, and their different expression level of gene and protein in pancreatic cancer cell lines were contrastly studied.

RESULTSNumber 3 and number 8 positive dots were identified as: voltage-dependent anion channel (VDAC3) and catechol-o-methyltransferase (COMT). RT-PCR, real-time PCR and Western blot showed that gene and protein of VDAC3 and COMT were expressed in the pancreatic cancer cell line SW1990, AsPc and P3 respectively.

CONCLUSIONVDAC3 and COMT might be the candidate immunogenic membrane antigens of human pancreatic cancer, and their gene and protein are differently expressed in the pancreatic cancer cell line SW1990, AsPc and P3.

Antigens, Neoplasm ; analysis ; Cell Line, Tumor ; Humans ; Pancreatic Neoplasms ; immunology ; Proteomics

The aim of this study was to apply a new type piezo-electric immunosensor in immunology classification for acute leukemia. Based on the plasma-polymerized film and the nanogold particles self-assembly technology, a new type piezo-electric immunosensor was firstly developed to absorb and fix the monoclonal antibodies for detecting leukemia cell antigens in patient blood samples. The results showed that the positive rate of detection with this method was quite close to that with the fluoroimmunoassay which are commonly used in clinic work. There was no significant difference between the positive rates (chi(2) = 3.4, P > 0.05). It is concluded that this piezo-electric immunosensor detection has many advantages, such as convenient operation, real time operation and control, qualitative detection, and less cost. This method can be used for acute leukemia immunology classification.
Acute Disease ; Antigens, Neoplasm ; analysis ; Biosensing Techniques ; methods ; Fluoroimmunoassay ; Humans ; Leukemia ; classification ; immunology

We describe a newly-made murine monoclonal antibody to the common acute lymphoblastic leukemia antigen (CALLA), named SHB-10. The antigen detected by SHB-10 has a molecular weight of about 105 kDa. This antibody is very similar to that of conventional anti-CD10 Ab on indirect flowcytometric analysis using lymphoid malignant cell lines and peripheral lymphocytes of acute lymphoblastic leukemia (ALL) patients. The binding of anti-CD10 to Daudi cell and peripheral lymphocytes of ALL patients is blocked by SHB-10. Thus this monoclonal antibody is thought to detect the CALLA. The distribution of antigen detected by SHB-10 on several cell lines of neuroectodermal tumor and lymphoid malignancy was analysed and a slight difference in their cell surface expression is observed when compared with that by conventional anti-CD10. Further biochemical analysis is now under way for a better characterization of this antigen.
Animals ; Antibodies, Monoclonal/*immunology ; Antigens, Differentiation/*analysis/immunology ; Antigens, Neoplasm/*analysis/immunology ; Flow Cytometry ; Humans ; Immunoglobulin Isotypes/analysis ; Mice ; Mice, Inbred BALB C ; Neoplasms/*immunology ; Neprilysin ; Tumor Cells, Cultured ; Tumor Markers, Biological/*analysis

Prostate stem cell antigen (PSCA) is a cell surface antigen expressed in normal prostate and overexpressed in cancers associated with prostate, bladder and pancreas. The sensitivity of PSCA labeling is higher than PSA in prostate cancer. PSCA can be used in the preparation of protein vaccine and nucleic acid vaccine. Further studies are required to confirm its safety and efficacy as a diagnostic means.
Antigens, Neoplasm ; GPI-Linked Proteins ; Humans ; Immunotherapy ; Male ; Membrane Glycoproteins ; analysis ; genetics ; immunology ; Neoplasm Proteins ; analysis ; genetics ; immunology ; Pancreatic Neoplasms ; diagnosis ; Prostatic Neoplasms ; diagnosis ; Urinary Bladder Neoplasms ; diagnosis

HLA expression is altered in a large variety of human cancers. We performed immunohistochemical staining on tissues from normal, preinvasive, invasive and metastatic cervical cancer tissues using anti-HLA class I or class II antibody. In tissues from normal squamous epithelium, carcinoma in situ (CIS) and microinvasive carcinoma (MIC), the expressions of HLA-B, C heavy chains and class II heavy chain were significantly decreased as disease progressed. When the expression patterns were compared between primary and metastatic squamous cell carcinoma (SCC) lesions, statistically significant down-regulation of HLA class I and class II antigen in metastatic lesions was observed. The rates of HLA-B, C heavy chains and class II heavy chain expressions were all significantly down-regulated compared to the down-regulation rate of class I beta2-microglobulin (beta2m) in invasive squamous lesions, and the expressions of class II heavy chain in metastatic lesions was decreased further than that in primary lesions. Unlike SCC, the degree of HLA class I and class II loss was not evident as disease progressed in early stage of adenocarcinoma. In invasive adenocarcinoma lesions, only the expression of HLA-B, C heavy chains was decreased and no differences were seen in HLA-B, C heavy chain expression patterns between primary and metastatic lesions. These results suggest that alterations of HLA class I and II expressions seem to occur at a particular step in cervical cancer development and depend on tissue types: when the tumor becomes invasive and starts to metastasize.
Antibodies, Monoclonal ; Carcinoma in Situ/immunology/pathology/physiopathology ; Carcinoma, Squamous Cell/immunology/pathology/physiopathology ; Cervix Neoplasms/*immunology/pathology/physiopathology ; Disease Progression ; Female ; Genes, MHC Class I ; Genes, MHC Class II ; HLA Antigens/*analysis ; HLA-B Antigens/analysis ; Histocompatibility Antigens Class I/*analysis ; Histocompatibility Antigens Class II/*analysis ; Human ; Immunohistochemistry ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Support, Non-U.S. Gov't