Bromodeoxyuridine, an analogue of thymidine, can be detected by means of monoclonal antibodies and utilized as a marker of the S-phase of the cell cycle. In vitro immunohistochemical application of the BrdU/anti-BrdU-MAb method permits a quantitative assessment of the proliferative activity of a tissue as well as the direct location of the actively replicating cells in histological sections. In this paper, a method for the detection of the labeling index of S-phase cells in normal and neoplastic tissues with in vitro BrdU labeling and standard immunohistochemical techniques using anti-BrdU-MAb and avidin-biotin peroxidase complex is described. We have employed this method in 47 human solid tumor samples, including squamous cell carcinomas of head and neck and cervix uteri, adenocarcinomas and malignant lymphomas, and also evaluated the possible application of the BrdU labeling index to estimate the cycling S-phase cells in neoplastic cell populations. In our data, the in vitro labeling index varied greatly in an individual case (3.56-29.2%) and from an area to an area within the same case. Squamous cell carcinomas of the head and neck showed higher LI than those of the cervix uteri. A case of metastatic carcinoma to the lung from ductal carcinoma of the breast had the highest LI (29.2%), in contrast to the low LI (3.6%) in the primary ductal carcinoma of breast.
Adenocarcinoma/immunology/pathology ; Antibodies, Monoclonal/*immunology ; Breast Neoplasms/immunology/pathology ; Bromodeoxyuridine/*immunology ; Carcinoma, Squamous Cell/immunology/pathology ; Cell Nucleus/immunology/*physiology ; Head and Neck Neoplasms/immunology/pathology ; Humans ; Immunohistochemistry ; *Interphase ; Lymphoma/immunology/pathology ; Neoplasms/immunology/*pathology

Antibodies, Monoclonal ; metabolism ; Antibodies, Monoclonal, Murine-Derived ; Antigens, CD34 ; metabolism ; Blood Vessels ; immunology ; pathology ; Breast Neoplasms ; immunology ; pathology ; Esophageal Neoplasms ; immunology ; pathology ; Female ; Humans ; Immunohistochemistry ; methods ; Lymphatic Vessels ; immunology ; pathology ; Neoplasm Invasiveness ; Stomach Neoplasms ; immunology ; pathology

Animals ; Antigens, Neoplasm ; biosynthesis ; genetics ; immunology ; Apoptosis ; physiology ; Breast Neoplasms ; immunology ; metabolism ; CD3 Complex ; immunology ; Female ; Humans ; Killer Cells, Natural ; pathology ; Neoplasms ; immunology ; metabolism ; Receptors, Antigen, T-Cell ; immunology ; Stomach Neoplasms ; immunology ; metabolism ; Uterine Cervical Neoplasms ; immunology ; metabolism

Clinical application of staphylococcal enterotoxin C2 (SEC2) was restricted during the cure of malignant tumor due to its side-effects. The aim of this study was to obtain SEC2 mutant, preserving the important functional sites responsible for the T-cell stimulatory activities but removing the sites responsible for emetic activity, through truncation of SEC2. It would efficiently solve the question of SEC2 side-effect. According to the results of methyl thiazol tetrazolium (MTT) assay in vitro, novel truncated SEC2 mutant (NSM) efficiently stimulated T-cell proliferation and inhibited the growth of such tumor cells as human colorectal cancer cells (Cx-1) and human breast cancer cells (MCF-7) in vitro. Activities of T cell stimulating and anti-tumor of NSM were similar to those of SEC2. According to results of animal experiments, the mutant no longer induced emetic response even if the dose was a 10-fold excess of the amount of SEC2 required. And also, NSM obviously inhibited the tumor growth in tumor-bearing mice. Therefore, we obtained novel truncated staphylococcal enterotoxin C2 mutant, which could efficiently inhibit the growth of tumor cells. It will become novel anti-tumor agents with the lowest side-effects and best treatment effects in clinic.
Animals ; Antineoplastic Agents ; adverse effects ; pharmacology ; Breast Neoplasms ; immunology ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Colorectal Neoplasms ; immunology ; pathology ; Enterotoxins ; genetics ; immunology ; Humans ; Mice ; Mutant Proteins ; immunology ; Staphylococcus aureus ; immunology ; Superantigens ; immunology ; T-Lymphocytes ; immunology ; Vomiting ; prevention & control

OBJECTIVETo evaluate the immunophenotype conversion of fibroblasts and its clinical significance in the process of breast tumor stromal remodeling.

METHODSCD34, FAP-α, p63 and a-SMA were detected by immunohistochemistry in 273 breast biopsies, including 60 normal breast tissues, 46 atypical ductal hyperplasia (ADH), 60 ductal carcinoma in situ (DCIS), 47 DCIS microinvasive carcinoma (DCIS-MI) and 60 invasive ductal carcinoma (IDC).

RESULTSThe positive expression rates of CD34, FAP-α and α-SMA in the stromal fibroblasts of normal breast tissues were 93.3%, 6.7% and 18.3%, respectively. Those in the stromal fibroblasts of ADH tissues were 95.7%, 4.3% and 10.9%, respectively. Those in the stromal fibroblasts of DCIS tissues were 95.0%, 8.3% and 15.0%, respectively. Those in the IDC tissues were 35.0%, 85.0% and 93.3%, respectively. The expressions of CD34, α-SMA and FAP-α in the stromal fibroblasts of normal, ASH and DCIS breast tissues did not show significant differences (χ(2) = 1.142, P = 0.896). The main immunophenotype of stromal fibroblasts in the tumor-host interface at the invasive front of ADH and DCIS lesions was CD34(+)α-SMA(+)FAP-α(+). There were statistically significant differences in the expression of CD34, α-SMA and FAP-α between IDC and ADH, DCIS and normal breast tissues (χ(2) = 8.351, P < 0.001). The immunophenotype of stromal fibroblasts in the IDC and DCIS-MI breast tissues was CD34(-) α-SMA(+) FAP-α(+).

CONCLUSIONSImmunophenotype conversion from CD34(+) α-SMA(-) FAP-α(-) to CD34(-) α-SMA(+)FAP-α(+) may be a sensitive indicator to judge whether DCIS has microinvasion. Detection of the immunophenotype conversion of stromal fibroblasts may be helpful to determine the presence of microinvasion, and to improve the diagnostic accuracy rate of DCIS.

Breast ; Breast Neoplasms ; immunology ; pathology ; Carcinoma in Situ ; Carcinoma, Ductal, Breast ; Carcinoma, Intraductal, Noninfiltrating ; Fibroblasts ; immunology ; Gelatinases ; metabolism ; Humans ; Hyperplasia ; Immunohistochemistry ; Immunophenotyping ; Membrane Proteins ; metabolism ; Serine Endopeptidases ; metabolism

OBJECTIVETo study the lymphocyte subsets of sentinel lymph nodes (SLN) in breast cancer patients.

METHODSFlow cytometry was used to analyze the markers on the surface of lymphocytes such as CD3, CD4, CD8, CD16 and CD19 in the sentinel lymph node of breast cancer.

RESULTSWhen lymph node metastasis did not occur, there was no significant difference in the number of CD3(+) T, CD4(+) T, CD8(+) T, CD16 NK and CD19 B cells between SLN cells and non-SLN cells. With lymph node metastasis, the proportion of CD4(+) and CD8(+) T cells was significantly changed, CD8(+) T cells (66.15 +/- 5.97) were the predominant cells instead of CD4(+) T cells (69.07 +/- 5.02), whereas no significant difference in CD3(+) T, CD16 NK and CD19 B cells.

CONCLUSIONThe CD4(+) to CD8(+) T cell ratio in sentinel lymph nodes with metastasis is reversed in breast cancer patients. This might results from changes in microenvironment due to tumor cell invasion.

Breast Neoplasms ; immunology ; pathology ; CD4-CD8 Ratio ; CD4-Positive T-Lymphocytes ; pathology ; CD8-Positive T-Lymphocytes ; pathology ; Carcinoma, Ductal, Breast ; immunology ; pathology ; Female ; Humans ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Lymphocyte Subsets ; immunology ; Sentinel Lymph Node Biopsy

OBJECTIVETo study the expression of a glycoprotein of plant origin in normal, benign and malignant breast tissues.

METHODSExpression of a plant glycoprotein was examined in 5 samples of normal breast tissues, 20 fibro-adenoma and 136 breast cancer by SABC immunohistochemical staining and the results were analyzed by SPSS statistics software.

RESULTSNo positive staining was detected in normal breast tissues (0/5). Weak staining was observed in 4 of 20 (20.0%) breast fibro-adenoma. Positive staining was demonstrated in 116 out of 136 (85.3%) breast cancer specimens. The differences were statistically significant. The expression of plant-associated human cancer antigen was related to pathological grade (P < 0.05), tissue invasiveness (P < 0.01) and recurrence (P < 0.05), but not to patients' age, tumor size and c-erbB-2 expression.

CONCLUSIONThe plant glycoprotein studied may be a human cancer-associated antigen which might be a potential marker of breast cancer.

Adenocarcinoma, Mucinous ; immunology ; pathology ; Adult ; Aged ; Antigens, Neoplasm ; metabolism ; Biomarkers, Tumor ; metabolism ; Breast ; immunology ; Breast Neoplasms ; immunology ; pathology ; Carcinoma, Ductal, Breast ; immunology ; pathology ; Carcinoma, Intraductal, Noninfiltrating ; immunology ; pathology ; Female ; Fibroadenoma ; immunology ; pathology ; Humans ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Recurrence, Local ; metabolism ; Plants ; immunology ; Receptor, ErbB-2 ; metabolism

OBJECTIVETo develop and identify the monoclonal antibodies (mAb) against c-erbB-2 p185 intracellular domain for detection of c-erbB-2 protein overexpression in breast tumor cells.

METHODSBALB/C mice were immunized with a synthesized p185 peptide of intracellular domain. The biological characteristics and immunoactivities were identified by different techniques.

RESULTSThree hybridoma cell strains secreting mAbs to c-erbB-2 protein were established and one of the mAbs, No. 035-E61 was tested for c-erbB-2 protein immunostaining on 39 breast carcinoma sections, 30 breast fibroadenoma sections and 16 sections from various normal organs. The results showed that the positive detection frequency of protein expression was 26% (10/39) in breast cancer, none (0/30) in fibroadenoma and 3 sections from normal organs were positively stained. The consistency between the 035-E61 and the DAKO reagent approved by FDA was 74%.

CONCLUSIONSNo. 035-E61 mAb can specifically recognize the p185 intracellular domain and may be useful in guiding clinical Herceptin treatment.

Animals ; Antibodies, Monoclonal ; immunology ; Antibody Specificity ; Binding Sites ; immunology ; Breast Neoplasms ; immunology ; pathology ; Female ; Fibroadenoma ; immunology ; pathology ; Humans ; Immunoglobulin G ; immunology ; Immunohistochemistry ; Mice ; Mice, Inbred BALB C ; Receptor, ErbB-2 ; immunology ; Tumor Cells, Cultured

It has been well established that immune surveillance plays critical roles in preventing the occurrence and progression of tumor. More and more evidence in recent years showed the host anti-tumor immune responses also play important roles in the chemotherapy and radiotherapy of cancers. Our previous study found that tumor- targeting therapy of anti-HER2/neu mAb is mediated by CD8(+) T cell responses. However, we found here that enhancement of CD8(+) T cell responses by combination therapy with IL-15R/IL-15 fusion protein or anti-CD40, which are strong stimultors for T cell responses, failed to promote the tumor therapeutic effects of anti-HER2/neu mAb. Analysis of tumor microenviornment showed that tumor tissues were heavily infiltrated with the immunosuppressive macrophages and most tumor infiltrating T cells, especially CD8(+) T cells, expressed high level of inhibitory co-signaling receptor PD-1. These data suggest that tumor microenvironment is dominated by the immunosuppressive strategies, which thwart anti-tumor immune responses. Therefore, the successful tumor therapy should be the removal of inhibitory signals in the tumor microenvironment in combination with other therapeutic strategies.
Animals ; Antibodies, Monoclonal ; immunology ; therapeutic use ; Breast Neoplasms ; drug therapy ; immunology ; pathology ; Cell Line, Tumor ; Female ; Humans ; Immune Tolerance ; immunology ; Mice ; Mice, Inbred BALB C ; Neoplasm Transplantation ; Receptor, ErbB-2 ; immunology ; Tumor Microenvironment ; immunology

OBJECTIVETo investigate the expression of indoleamine 2, 3-dioxygenase (IDO) in breast cancer and its correlation with clinicopathologic factors and prognosis.

METHODSThe expression of IDO, CD31, CD105 proteins in 40 specimens of breast cancer were assessed by immunohistochemistry.

RESULTSThe overexpression rate of IDO in breast cancer was 67.5% (27/40), and expression of IDO was closely associated with clinical stage and lymph nodes metastasis. The disease-free survival rate in patients with IDO overexpression was not significantly lower than that in patients with negative or low expression of IDO (P > 0.05). Moreover, the expression of IDO was positively correlated with CD105-labeled microvessel density (r = 0.659, P < 0.05).

CONCLUSIONSExpression of IDO is associated with clinical stage and lymph nodes metastasis, and microvessel densitty. IDO expression may promote the growth and metastasis of breast cancer, probably via the increased agiogenesis. A larger sample study is needed to verify whether the prognosis of beast cancer is significantly correlated with IDO expression.

Adenocarcinoma ; enzymology ; immunology ; pathology ; Adult ; Aged ; Antigens, CD ; metabolism ; Breast Neoplasms ; enzymology ; immunology ; pathology ; Carcinoma, Ductal, Breast ; enzymology ; immunology ; pathology ; Carcinoma, Medullary ; enzymology ; immunology ; pathology ; Disease-Free Survival ; Endoglin ; Female ; Follow-Up Studies ; Humans ; Immunohistochemistry ; Indoleamine-Pyrrole 2,3,-Dioxygenase ; metabolism ; Lymphatic Metastasis ; Microvessels ; enzymology ; immunology ; Middle Aged ; Neoplasm Staging ; Platelet Endothelial Cell Adhesion Molecule-1 ; metabolism ; Receptors, Cell Surface ; metabolism ; Survival Rate