Relationship of monoclonal antibody binding to estrogen and progesterone receptor content in breast cancer

Three monoclonal antibodies--H59, H71, and H72--which react with human breast cancers have been developed using the estrogen-dependent human breast cancer cell line, ZR-75-1, as the immunogen. H59 bound only to estrogen receptor-positive, estrogen-regulated breast cancer cells in culture, whereas H71 and H72 bound breast cancer cells irrespective of the estrogen receptor content. All three antibodies have minimal cross-reactivity with non-breast tissue culture cell lines. The three antigens appear to be glycoproteins located on the cell surface. H59 and H72 antigens bound preferentially to the apical surface of duct cells and may be secreted; H71 antigen demonstrated no evidence of an apical orientation or secretion. The binding of the antibodies to fixed cryosections from 152 breast cancer and 111 benign breast disease specimens has been evaluated using a radioimmunoassay. Eighty-five % of breast cancer and almost 100% of benign disease specimens were bound by at least one antibody. H59 bound 39%, H71 bound 51%, and H72 bound 65% of cancer specimens. Estrogen receptor and progesterone receptor analyses were obtained on 141 specimens. H59 bound almost exclusively to tumor specimens which contained estrogen and/or progesterone receptor, but not to all receptor-positive tumors. Therefore, the H59 antigen appeared to be present on a subset of estrogen receptor-positive tumors. Considering that it bound only to estrogen-regulated cells in culture, the antigen may be estrogen regulated, and its presence may predict a response to hormone therapy. H71 and H72 recognized cell surface differentiation antigens but bound tumor specimens regardless of the receptor content. These antibodies may be useful as independent variables for predicting response to therapy and prognosis of patients with breast cancer.     

Prognosis in breast cancer utilizing histologic characteristics of the primary tumor.

A study was made of the inter-relationships and prognostic significance of structural characteristics found in primary breast cancers and their associated axillary lymph nodes. The prognostically favorable characteristics included the following. For the primary tumor: nuclear differentiation of the cancer cells, diffuse lymphoid cell infiltrations (LI) and perivenous lymphoid cell infiltrations (PVI). For the axillary lymph nodes: sinus histiocytosis (SH). Perivenous lymphoid cell infiltrations (PVI) in the primary tumor are found to be as important a prognostic factor as SH in the axillary lymph nodes; these two characteristics are found to be positively associated. Evaluation of the nuclear grade (NG), LI, and PVI in the primary tumor allows for the definition of association with cancer cells having a low (anaplastic) nuclear grade, and to be positively associated with follicular hyperplasia (FH) in the lymph nodes. We also found a positive association between the cellular responses to areas of in situ carcinoma and the cellular responses to accompanying invasive breast breast cancer tissue. They also provide a prognostic system for classifying breast cancer patients on the basis of the microscopic characteristics of the primary tumor and surrounding breast tissue. The latter system should be of value in comparing the therapeutic benefits of various treatments.     

Increased expression of Thomsen-Friedenreich antigens during tumor progression in breast cancer patients

Paraffin-embedded tissue sections of primary tumors and lymph node metastases of 80 breast cancer patients were tested for the expression of Thomsen-Friedenreich (TF) antigens with the aid of a monoclonal IgM antibody (49H8) highly specific for phenyl-beta-galactoside. TF antigens were not expressed in 16 different normal tissues with the exception of some structures in the kidney. In tumor cells, two types of antigen expression were found; namely, cryptic and exposed. From stage T1/No to stages T2-4/N1,2 the number of cases expressing high amounts of TF antigens increased from 9% (2/22) to 22% (4/18) while the percentage of patients with low intensity of antibody binding was reduced from 59% (13/22) to 39% (7/18). The total amount of TF-positive primary tumors at stages T2-4/No increased from 42% (8/19) to 69% (18/26) when lymph nodes were infiltrated (T2-4/N1,2). At this stage 80% (21/26) of the patients with lymph node infiltration carried TF antigens in the nodes. The distribution of antigens was heterogeneous among the tumor cells and was expressed mainly in an apical or luminal position. The increased expression of antigens was attributed to exposed TF antigens, while cryptic antigens remained constant. When primary tumors expressed exposed TF antigens, the corresponding lymph nodes also contained exposed antigen. The same was true for the cryptic antigen. The data demonstrate an increase in the intensity of TF antigen expression during tumor progression and a spread of TF-positive tumor cells into the axillary lymph nodes with an increasing number of breast cancer patients being TF-positive at this stage.     

Isolation of native human monoclonal autoantibodies to breast cancer

Using a unique fusion partner cell line, MFP-2, and B-lymphocytes from breast cancer patients, we developed a set of fully human monoclonal antibodies (MAbs) that bind with high specificity and sensitivity to breast cancer cells. Immunofluorescent staining of normal tissues, primary tumors, and metastatic lymph nodes demonstrates that these antibodies are specific for breast cancer of autologous and allogeneic origin. We have also determined that many of the antibodies selected based on specific binding to breast cancer cells and tissue also bind prostate cancer cells and tissue with high specificity and sensitivity. The targets of these antibodies have been localized to the cytoplasm and membrane. Biological assays for internalization and cytotoxicity demonstrated the ability of three antibodies to rapidly internalize. Our study demonstrates that isolation of native human MAbs from the natural antibody repertoire, targeted to cancer cells, is feasible and may provide a source of tools for immunotherapy.     

Differential Expression of RAGE,EGFR and Ki-67 in Primary Tumors and Lymph Node Deposits of Breast Carcinoma

Background: Breast cancer is a complex disease that results from the inheritance of a number of susceptible genes.Intensive search wok was conducted world-wide on molecular bases of breast cancer in order to achieve the besttherapeutic modalities; however, breast cancer still remains a challengeable task. It is very important to determine ifthe biological parameters in metastatic regional lymph nodes are similar to that in the primary breast cancer becausetherapy is indicated for patients with synchronous metastatic regional lymph nodes of breast cancer. Difference intherapeutic response in cases of breast cancer may be assumed partially to variability in the biological behavior of tumortissue in primary breast cancer and lymph node metastasis. Aim: Our aim is to evaluate any variability in the expression ofthree types of tissue markers in both the primary breast tumors and corresponding axillary lymph nodes in order toexpect the targeted therapeutic effect on both sites. Material and Methods: Three markers from different categories;RAGE, EGFR and Ki-67 were immunohistochemicalyl studied for their expression in biopsy specimens from primarybreast tumors and their corresponding axillary lymph nodes. Results: There was a statistically significant difference inthe expression of these markers between benign and malignant breast lesions.Although we found some differences inthe expression of the three studied markers between primary breast cancer and corresponding axillary lymph nodes, yetthese variations were mostly not statistically significant. Conclusion: Our findings support the validity of anti-RAGEand anti-EGFR therapy for treatment of both primary and nodal metastatic breast cancer in immunopositive cases.     

Cigarette smoking and breast cancer risk: update of a prospective cohort study

SummaryIntroduction Ultrasound (US) preoperative examination of the axillary lymph nodes combined with the fine needle aspiration biopsy (FNAB) is often used in order to reduce the number of sentinel lymph node (SLN) biopsy procedures in clinically node negative breast cancer patients. The pathohistological characteristics of the ultrasonically negative axillary lymph nodes in clinically negative axillary lymph nodes are not known. The aim of our study was to compare the pathohistological characteristics of ultrasonically uninvolved axillary lymph nodes (US group) versus clinically uninvolved axillary lymph nodes (non-US group) in SLN biopsy candidates.Methods We included 658 patients after SLN biopsy; 286 patients in the US group and 372 in the non-US group. The pathohistological characteristics of axillary lymph nodes were evaluated by univariate analysis and logistic regression.Results In the univariate analysis, the proportion of macrometastastic SLN, total number of metastatic lymph nodes per patient, proportion of nonsentinel lymph node (NSLN) metastases and proportion of NSLN macrometastases were found to be lower in the US group compared to the non-US group. In the logistic regression model, only US of the axilla (p=0.010; OR: 0.57) and tumor size were significant predictors for the presence of SLN macrometastases or macrometastatic NSLN (p<0.001; OR: 0.23).Conclusion The patients with US negative axillary lymph nodes form a distinct subgroup of early breast cancer patients having a significantly lower tumor burden in the axillary lymph nodes compared to those with only clinically negative axillary lymph nodes.     

Is dissection of the internerve tissue during axillary lymphadenectomy for breast cancer necessary?

AIMS: The study evaluates the necessity of dissecting the tissue between the long thoracic and thoracodorsal nerves (internerve tissue) during axillary dissection in breast cancer surgery. By reviewing the lymph node yield and the metastatic rate in the internerve tissue, we examine whether the internerve tissue could be left in situ to minimize the risk of nerve injury. METHODS: A prospective study was conducted on 30 consecutive women undergoing axillary lymphadenectomy for breast cancer. The internerve tissue remaining was excised separately after a routine axillary dissection and was examined by the same pathologist. RESULTS: Twenty (67%) of 30 internerve specimens contained lymph nodes; the internerve nodes were positive for carcinoma in three cases (10%). In one case the lymph node in the internerve tissue was the only metastatic node in the axilla. CONCLUSIONS: There is a significant incidence of lymph nodes (67%) and axillary node metastases (10%) in the tissue lying between the long thoracic and thoracodorsal nerves. Therefore excision of this internerve tissue is strongly recommended in order to optimize decision making regarding adjuvant treatment and oucome in women with operable breast cancer.     

Survival with mammary cancer related to the interaction of germinal center hyperplasia and sinus histiocytosis in axillary and internal mammary lymph nodes.

In a review of the histologic sections of axillary and internal mammary lymph nodes removed during surgery for invasive ductal carcinoma of the breast, we found that 16 of 17 patients in whom sinus histiocytosis was the dominant lymphoid proliferative reaction are alive with no evidence of cancer 5 or more years after operation. In contrast, 5 of 6 patients in whom germinal center hyperplasia was the only significant reaction found died of cancer in less than 5 years. Patients with both sinus histiocytosis and germinal center hyperplasia in significant amounts had survival that was intermediate; 17 of 25 of these patients are currently alive and apparently free of cancer. In addition, 5 of 6 patients in whom no evidence was found of any lymphoid proliferative reaction and 3 of 3 patients with diffuse cortical hyperplasia in their axillary lymph nodes died of cancer in less than 5 years. Germinal center hyperplasia was associated with nodal metastases anatomically in individual lymph nodes and statistically in the series of cases. The internal mammary lymph nodes of most cases showed less proliferative reaction to tumor than the axillary lymph nodes. The pattern of proliferative reactions in lymph nodes and its correlation with survival after surgery suggest that different immune reactions may either suppress or enhance the growth of carcinoma of the breast.     

纳米炭对乳腺癌腋窝淋巴结示踪效果及其安全性的初步研究

目的探讨国产纳米炭混悬液对腋窝淋巴结的示踪效果及其安全性。方法2008年5月至2009年9月间对21例乳腺癌患者在术前采用纳米炭进行淋巴结示踪。患者年龄30～65岁,平均45．3岁。术前24～72h于乳晕周围分4点皮下均匀注射纳米炭混悬液共1ml。手术方式采用改良根治术5例,腔镜下乳房皮下腺体切除、腋窝淋巴结清除加假体植入术12例,乳腺癌局部扩大切除加腔镜腋窝淋巴结清除术4例。术后观察腋窝淋巴结的黑染情况并送病理检查。结果21例患者分别检出淋巴结14～32枚,平均每例21．5枚,共452枚。肉眼下黑染淋巴结共435枚,黑染率为96．3％（435／452）,其中明显黑染率为85．4％（386／452）。腋窝淋巴结无转移6例,有转移15例。有转移的淋巴结共45枚,均为明显黑染淋巴结,转移淋巴结黑染率为100％。经病理证实,未黑染的淋巴结均未出现癌转移。所有患者经8个月至2年的随访,均未出现复发转移及明显肝肾功能异常。有纳米炭残留的局部组织在术后1年行活组织检查,见乳腺组织间隙有较多的纳米炭颗粒沉集,但未见明显的炎症反应或组织变性。结论纳米炭混悬液经乳晕周围皮下注射后24～72h行腋窝淋巴结清除可达到良好的淋巴结示踪效果,有效避免转移淋巴结的漏检。残留体内的纳米炭无明显的毒副反应。纳米炭是一种安全可靠的腋窝淋巴结示踪剂。     

Prognostic significance of mRNA-encoding estrogen receptor and epithelial growth factor receptor in breast carcinoma progression into lymph nodes: 1. Estrogen receptor encoding mRNA

The main objective of this study was to differentiate between lymph nodes infiltrated by estrogen receptor-positive (ER+) and estrogen receptor-negative (ER-) breast carcinoma. Lymph nodes were obtained from 40 postmenopausal cancer patients, 10 from each disease stage. Six patients from each group had estrogen receptor-positive (BCaER+) and four estrogen receptor-negative (BCaER-) tumors. Both tumor-containing (T) and uninvolved (N) lymph nodes from the same patient were examined by the following parameters: magnitude of lymph node nucleic acid hybridization with cDNA probes from breast cancer MCF-7ER+ and MCF-7ER- cells; and binding capacity of 3H-estradiol, 125I-EGF, and 125I-PDGF binding and protein kinase C activities of the lymph nodes. Concomitant with the appearance of transformed cells, several events occur: Tumor cells induce stimulation of mononuclear cells and macrophages and evoke T- and B-cell proliferation, leading to the synthesis of tumor cell membrane-associated antibodies. In estrogen receptor-positive (ER+) breast carcinoma, estrogens and host hormonal modulatory mechanisms stimulate production and release of epithelial growth (EGF) and platelet-derived growth factors (PDGF). These factors are characterized by protein kinase C activities. There is infiltration of tumor cells into the lymph node and infiltration of leukocytes into the tumor site. In the lymph node, tumor progression depends on tumor cell proliferation rate and metastatic aggressiveness. The experiments described in this study document the changes that occur in lymph nodes, with differences between nodes infiltrated with BCaER+ and BCaER- breast carcinomas. Hybridization of 32P-cDNA from MCF-7ER+ cells with cellular RNA from BCaER+ involved (T) lymph nodes is greater than with cellular RNA from uninvolved (N) lymph nodes. The magnitude of hybridization correlated (P less than 0.005) with the disease stage.     

The impact of axillary lymphadenopathy on further treatment in Breast Cancer? A model for clinical staging

Clinical assessment is an important part of the breast cancer patients’ work-up, but it has low sensitivity and specificity. In a retrospective study, his-tological slides of axillary clearance specimens were used to model palpability of the axillary lymph nodes. Obvious nodes (enlarged and involving considerable amount of lymphatic and/or metastatic tissue) and nodes equal to or larger than 1 cm or 1.5 cm were counted and the slides were subsequently reviewed. The false positive and negative rates expected on the basis of the model ranged from 24 to 72% and from 10 to 38%, respectively. This model (also valid for intraoperative assessment of nodal status by palpation) documents the lack of specificity of clinical staging of the axilla. These results question the practice of excluding patients with palpable axillary lymph node enlargement from less radical staging procedures such as axillary sampling or sentinel node biopsy     

Interferon production in lymph nodes draining breast carcinoma and its augmentation by OK-432

In breast cancer patients on whom modified radical mastectomy is performed, relatively more of the regional lymph nodes draining the breast carcinoma remain in comparison with standard radical mastectomy. Therefore, investigation of the functions of lymph nodes draining breast carcinoma has become important. Lymphocyte subsets of 33 axillary lymph nodes from 19 breast cancer patients were analysed using flow cytometry. In axillary lymph nodes, both OKT-3(+) cells and OKT-8(+) cells were decreased in comparison with those in peripheral blood. However, the OKT 4/8 ratio was increased in axillary lymph nodes. These findings suggest that axillary lymph nodes are immunologically more functional against cancer spread than peripheral blood. OK-M1(+) cells, Leu-7(+) cells and Leu-11a(+) cells were decreased in axillary lymph nodes in comparison with peripheral blood. The ability of IFN production in axillary lymph nodes and peripheral blood was analysed using the cytopathic effect of VSV-sindbis virus. After 72 hours incubation, IFN production of axillary lymph nodes showed maximum titer. When lymph nodes were co-cultured with OK-432, IFN production of axillary lymph nodes was strongly augmented. IFN production of axillary lymph nodes draining breast carcinoma were increased in comparison with peripheral blood. Axillary lymph nodes draining breast carcinoma would thus seem to be important as cytokine-producing organs. IFN has been found to be an activator of NK cells, cytotoxic T cells and IL-2 production. Axillary lymph nodes may therefore play an important role against the spread of breast cancer.     

Tissue harmonic imaging sonography of the axillary lymph nodes: evaluation of response to neoadjuvant chemotherapy in breast cancer patients

In breast cancer patients, the number of surgically resected metastatic axillary lymph nodes has been considered to correlate with prognosis. Therefore, the lymph nodes' response to chemotherapy may provide a favorable prognosis by suggesting an appropriate regimen of post-operative chemotherapy. In the present study, we evaluated the therapeutic efficacy of neoadjuvant chemotherapy of the axillary lymph nodes of breast cancer patients, using tissue harmonic imaging ultrasonography. Thirty-three female patients with breast cancer >2 cm in the longest diameter were examined by tissue harmonic imaging both before and following neoadjuvant chemotherapy. Nodes were defined as metastasis positive or negative on the basis of Yang's criteria, at each ultrasonography examination. The ultrasonography findings were correlated with the histopathologic results of the surgically resected specimens. Patients with constantly positive nodes over two ultrasonography examinations had a higher pathological nodes positive rate (8/9, 88.9%) than those (3/10, 30.0%) whose lymph nodes shrunk from the first to second ultrasonography exam (p=0.02, Fisher's exact method). All patients with negative nodes at both ultrasonography examinations had negative pathological results. Thus, changes in consecutive tissue harmonic imaging findings may be used to accurately evaluate the response of axillary lymph nodes to neoadjuvant chemotherapy.     

Estrogen Receptor Mutations and Changes in Estrogen Receptor and Progesterone Receptor Protein Expression in Metastatic or Recurrent Breast Cancer

To investigate the frequency of estrogen receptor ( ER ) gene mutation in metastatic or recurrent breast cancer, metastatic lymph nodes or recurrent breast cancer tissue from 35 patients with ER-positive primary tumors were screened for mutations in the hormone-binding domain of the ER gene by sequence analysis. Four missense mutations, Val316Ile, Gly344Val, Ala430Val and Gly494Val, were identified in these lesions. Second, to clarify whether there is any disparity in hormone receptor status between primary and metastatic or recurrent tumors, we immunohistochemically studied 117 specimens including the above 35 specimens obtained from metastatic or recurrent breast cancer patients using monoclonal anti-ER and progesterone receptor (PgR) antibodies. Although hormone receptor status, especially ER, was highly maintained through disease progression, negative change in PgR expression at relapse (33%) was identified more frequently than in metastatic lymph nodes (6.7%). Therefore, it was suggested that development of PgR-negative phenotype might correlate with disease progression in some breast cancer patients. These results suggest that ER mutations in metastatic or recurrent breast cancer may be more frequent than in primary lesions, irrespective of high maintenance of ER protein expression through disease progression.     

术前淋巴化疗对乳腺癌复发转移的影响及其机制

背景与目的:有报道认为淋巴化疗可以增加消化道肿瘤引流区域淋巴结中抗癌药物的浓度,改善预后,关于乳腺癌淋巴化疗的研究尚少见报道。本研究探讨术前淋巴化疗对乳腺癌复发转移的影响及其作用机制。方法:将60例Ⅱ～Ⅲ期乳腺癌患者随机分为两组,淋巴化疗组40例,对照组20例。在改良根治术前72h,淋巴化疗组于肿瘤周围或瘤床注射表阿霉素-活性炭混悬液10mg;对照组注射表阿霉素水溶液10mg,注射部位和方法同淋巴化疗组。用末端转移酶标记法检测腋窝转移淋巴结癌细胞凋亡指数(apoptoticindex,AI);用免疫组化SP法检测Fas/Fas-L蛋白的表达。观察两组患者局部和全身反应,比较复发转移率。结果:淋巴化疗组转移癌细胞AI平均为(9.5±2.7)%,对照组平均(3.8±1.4)%,前者明显高于后者(P﹤0.01);淋巴化疗组Fas蛋白表达比对照组明显上升(P﹤0.05),而Fas-L蛋白表达无明显改变(P﹥0.05)。对照组中有5例患者出现注射部位皮肤红肿发热,两组均无与化疗有关的局部和全身不良反应。淋巴化疗组患者的2年复发转移率为10.34%,低于对照组的38.46%(P﹤0.05)。结论:乳腺癌术前应用活性炭-表阿霉素混悬液淋巴化疗,可能会降低乳腺癌的复发转移率,其作用可能是通过上调Fas蛋白表达、诱导腋窝转移淋巴结癌细胞凋亡实现的。     

Correlation of Bcl-2 and p53 expression in primary breast tumors and corresponding metastatic lymph nodes

BACKGROUND: The p53 tumor suppressor gene product participated in G1 cell cycle arrest or cell death. Loss of function was associated with poor outcome in patients with breast carcinoma. bcl-2 prevented apoptosis induced by c-myc or growth factor deprivation. High bcl-2 expression in breast tumor tissue specimens appears to be associated with favorable prognostic factors. However, Bcl-2 and p53 expression in primary tumor tissue specimens versus metastatic lymph node specimens in breast carcinoma has not been studied. The current study compared Bcl-2 and p53 expression in primary breast carcinoma tissue specimens with Bcl-2 and p53 expression in axillary lymph node specimens. METHODS: Primary breast tumor and corresponding axillary metastatic lymph node tissue specimens were obtained from 60 patients with breast carcinoma. They were evaluated for the presence of Bcl-2 and p53 expression by immunohistochemistry using standard methods. RESULTS: Bcl-2 expression in primary tumor tissue specimens (53%) was correlated with Bcl-2 expression in metastatic lymph node specimens (50 %; Pearson correlation = 0.656). p53 expression in primary tumor specimens (72%) was correlated with p53 expression in metastatic lymph node specimens (60 %; Pearson correlation = 0.800). A significant inverse correlation also was found between p53 and Bcl-2 expression in primary breast tumor tissue specimens (Pearson correlation = -0.310). CONCLUSIONS: The current study suggested that Bcl-2 and p53 expression in axillary metastatic lymph node specimens is correlated with Bcl-2 and p53 expression in the primary tumor tissue specimens. The prognostic and predictive value of Bcl-2 and p53 expression in axillary lymph node metastasis in patients with breast carcinoma needs to be further evaluated in larger trials with longer follow-up.     

Generation and immunohistological characterization of human monoclonal antibodies to mammary carcinoma cells

The purpose of this study was to identify human antibodies generated against autologous breast tumor cells by the host's immune response. Accordingly, lymphocytes from lymph nodes of seven different patients with metastatic breast carcinomas were immortalized by fusing them with a nonsecreting variant of murine myeloma cells. The screening for binding of antibodies to tumor cells was performed by indirect immunoperoxidase staining of paraffin-embedded tissue sections of the autologous tumor. The selected hybrid cells, after being cloned three times, were stable for the secretion of immunoglobulins for over 2 years. A total of 81 human immunoglobulin-producing clones was obtained from an initial 595 wells with hybrid growth. Nine of these clones produced immunoglobulin M, none of which showed detectable binding to tissue antigens in breast. Seventy-two clones produced immunoglobulin G monoclonal antibodies, and 15 of these showed preferential binding to breast carcinoma cells. Three of these immunoglobulin G monoclonal antibodies were subjected to detailed immunohistological evaluations. Using these antibodies at concentrations ranging from 10 to 100 ng/tissue section, the morphologically normal mammary epithelial cells could be discriminated from their malignant counterparts. The antibodies showed diffuse staining of cytoplasmic components in the malignant counterparts. Under these conditions, lymphocytes, erythrocytes, and stromal cells in breast tissues were unstained. The antibodies showed variable reactivity with malignant epithelial cells of colon and stomach, and with normal epithelial cells lining the renal tubules and sebaceous glands in skin. Antigenic heterogeneity of malignant mammary epithelial cells was revealed. The antibodies may have value in the characterization of tumor-associated antigens responsible for inducing autologous immune responses.     

The immunohistochemical detection of lymph node metastases from infiltrating lobular carcinoma of the breast

Immunological markers improve specificity and accuracy of cell detection, therefore it is important to evaluate their usefulness in improving standard histological procedures. This study investigates whether immunocytochemical techniques increase the accuracy of detection, in axillary lymph nodes, of metastatic cells from infiltrating breast lobular carcinoma (ILC). Fifty cases of ILC reported to be node-negative were selected. New serial sections were cut from a total of 767 lymph nodes, stained with H&E and tested in immunoperoxidase (ABC procedure) with a conventional anti-Epithelial Membrane Antigen (EMA) serum, with a monoclonal raised against human milk fat globule membranes (HMFG-2) and with a monoclonal against 54 kd keratin. Metastases were detected immunocytochemically in 12 cases (24%); in five of these cases metastatic cells were also visible in serial H&E sections. Monoclonals offered no evident advantage over anti-EMA conventional antiserum. Immunocytochemical positivity alone is not sufficient evidence for metastatic invasion since macrophages occasionally appear EMA- and HMFG-2-positive (probably because of secondary incorporation of the antigen), and so an improvement in the accuracy of breast cancer metastatic cell detection in axillary lymph nodes requires a combined histo-immunological approach.     

Heterogeneity in the expression of HLA and tumor-associated antigens by surgically removed and cultured breast carcinoma cells

Surgically removed normal and malignant mammary tissues and human breast carcinoma cell lines were tested in binding assays with monoclonal antibodies to HLA-A,B,C antigens, beta 2-microglobulin, HLA-DR antigens, and tumor-associated antigens; the latter included a Mr 280,000, a Mr 94,000, and a Mr 85,000 membrane-bound glycoprotein and a cytoplasmic antigen. HLA-A,B antigens, beta 2-microglobulin, HLA-DR antigens, and the cytoplasmic antigen are expressed by normal mammary cells. Their malignant transformation may be associated with quantitative changes in the expression of these antigens and with the appearance of Mr 94,000 and Mr 85,000 glycoproteins. The Mr 280,000 glycoprotein was detected on only one of the breast carcinoma cell lines tested. Analysis of primary tumors and autologous axillary lymph node metastasis from 13 patients has shown differences in the expression of all the antigens tested between primary and metastatic lesions.