Immunohistological patterns of myeloid antigens: tissue distribution of CD13, CD14, CD16, CD31, CD36, CD65, CD66 and CD67

Summary. A number of differentiation antigens on myeloid cells have been defined on the CD classification system by the four International Workshops on Human Leucocyte Differentiation Antigens. The distribution of eight of these antigens (CD13, CD14, CD16, CD31, CD36, CD65, CD66, CD67) have been studied in human tissues, with the aim of documenting their immunohistological patterns and their degree of myeloid restriction. CD13, the most widely distributed antigen, was found in skin, bile canaliculi, kidney and pancreas. CD14 was not restricted to monocytes and tissue macrophages, being also strongly expressed on dendritic reticulum cells. CD 16 was expressed on granulocytes and tissue macrophages (alveolar and Kupffer cells) and in the red pulp of the spleen. CD31 and CD36 gave a characteristic staining of vascular endothelium, corresponding to their identification as the platelet glycoproteins gp IIa and gp IV. Antibodies against the most recently defined myeloid antigens (CD65, CD66 and CD67) appeared to be more specific for myeloid differentiation than previously described 'myeloid antigens'.     

Value of monoclonal anti-CD22 (p135) antibodies for the detection of normal and neoplastic B lymphoid cells

Two monoclonal antibodies (To15 and 4KB128) specific for the B cell- associated CD22 antigen (135,000 mol wt) are described. On immunoenzymatic analysis of cryostat tissue sections, these antibodies strongly label both mantle zone and germinal center B lymphoid cells in secondary lymphoid follicles (and also scattered extrafollicular lymphoid cells) but are unreactive with other cell types (with the exception of weak reactivity with some epithelioid histiocytes). These reactions differ from those of monoclonal antibodies B1 and B2 (anti- CD20 and CD21) but are similar to those of the pan-B antibody B4 (anti- CD19). One of the anti-CD22 antibodies (To15) has been tested extensively by immunoenzymatic labeling on greater than 350 neoplastic lymphoid and hematological samples. The CD22 antigen was found in tissue sections in most B cell-derived neoplasms, the major exceptions being myeloma (all cases negative) and a small proportion of high-grade lymphoma (6% of cases negative). In cell smears, the antigen could be found on neoplastic cells in most B cell lymphoproliferative disorders, including common acute lymphoblastic leukemia (ALL) (90% positive) and B cell chronic lymphocytic leukemia (CLL) (89% positive). We conclude that anti-CD22 antibodies are of value for identification of human B cell lymphoproliferative disorders (especially when used in conjunction with anti-CD19 antibodies). Previous reports that the CD22 antigen is absent from many B cell neoplasms are probably due to its being expressed within the cytoplasm of immature B cells rather than on their surface.     

Immunophenotypic correlation between skin biopsy and peripheral blood findings in mycosis fungoides

In mycosis fungoides (MF) with blood involvement, T-cell immunophenotypes in skin and blood have not been compared. Our aim was to evaluate T-cell immunophenotypes in skin by immunohistochemical analysis and compare results with flow cytometric (FC) findings in blood. Of 20 patients with MF with blood involvement, the immunophenotype was discrepant in 11 (55%). Compared with FC findings in blood, immunohistochemical analysis of skin samples failed to detect partial deletion of CD2 (5/11 [45%]), CD3 (3/11 [27%]), and CD5 (3/11 [27%]) and overrepresented deletion of CD7 in 2 (18%) of 11 patients. In addition, CD8+ MF was missed by immunohistochemical analysis in 2 (18%) of 11 patients. Identical T-cell populations were demonstrated by T-cell gene polymerase chain reaction in skin and blood in 8 of the 11 patients who had a discrepant immunophenotype. Awareness of the limitations of immunohistochemical analysis of skin samples is of practical value for pathologists interpreting skin biopsies in MF patients. In addition, our findings suggest CD8+ MF to be more common than previously reported.     

LC3A-Positive Light Microscopy Detected Patterns of Autophagy and Prognosis in Operable Breast Carcinomas

Autophagy is a self-degradation mechanism by which cells recycle their own cytoplasmic constituents and dispose of excess or defective organelles after starvation and oxygen deprivation. An antibody to the microtubule-associated protein 1 light chain 3 (LC3A), recognizing both the soluble (LC3A-I) and the membrane-bound form (LC3A-II) of the protein, was used to detect autophagic activity in 102 breast carcinomas. Three distinct patterns were recognized: (1) diffuse cytoplasmic, (2) cytoplasmic/juxta-nuclear, and (3) “stone-like” pattern – dense, rounded, amorphous structures, 5 μm on average, typically enclosed within cytoplasmic vacuoles. The diffuse cytoplasmic pattern showed a direct association with estrogen and progesterone receptor expression. The juxta-nuclear pattern indicated a similar association with hormone receptors, an inverse association with tumor size, and a favorable prognosis. By contrast, an increased number of stone-like structures, probably representing an excessive autophagic response, was related to high-grade tumors and a less favorable outcome. Interestingly, 60 additional epithelial tumors of nonbreast origin disclosed identical autophagic patterns, and so did MDA231 breast cancer xenografts and HCT116 colon tumor spheroids (also analyzed by electron microscopy). Moreover, MCF-7 human breast cancer cell lines confirmed induction of LC3A by anoxia and Thapsigargin. It is concluded that autophagy can be readily recognized in breast carcinomas by light microscopy, after immunohistochemical staining with LC3A, but the significance of the various patterns expressed would need further evaluation.Cancer cells survive the adverse conditions of the extracellular milieu (i.e., hypoxia, nutrient deprivation, and reduced growth factors) through angiogenesis and anaerobic glycolysis.¹ Yet rapidly proliferating malignant neoplasms, having high metabolic demands, are not sufficiently supplied by these processes.^2,3,4,5,6 An alternative metabolic pathway for providing energy, when both oxygen and glucose are depleted, is autophagy—a self-degradation mechanism by which cells recycle their own cytoplasmic constituents and dispose of excess or defective organelles resulting in protein and ATP synthesis.^7,8,9 However, autophagy when prolonged can also cause cell death, but how the outcome is determined and the relationship of autophagy to clinical outcome is, at present, poorly understood.Angiogenesis and anaerobic glycolysis (i.e., the shifting from oxidative phosphorylation to anaerobic metabolism) have both been studied extensively in relation to malignant disease, including breast cancer.^{10,11,12,13,14,15} The activation of antiapoptotic pathways, as a complementary cell surviving mechanism, has also been considered in a number of studies.^16,17 By contrast, the phenomenon of autophagy only recently received attention for its biological significance in human malignancies. It is now known, for example, that under suboptimal microenvironmental conditions, cytoplasmic constituents are first entrapped by a membrane sac, called the isolation membrane, which subsequently closes to form double membrane structures, called autophagosomes or autophagic vacuoles. These, and specifically their outer membrane, are fused with lysosomes producing autolysosomes.^8,18 Lysosomal hydrolases degrade the cytoplasmic contents of the autophagosome, together with its inner membrane,^8,18 and the resulting macromolecules are recycled. Failure to accomplish this self-degradation process, probably because of overload of cytoplasmic components, enzyme exhaustion, or lysosomal enzyme defects, leads to accumulation of debris and massive destruction of tumor cells.^18,19,20,21Increased new blood vessel formation is associated with rapidly advancing tumors.^10,12,13 Equally, high levels of hypoxia are indicative of tumor aggressiveness.^11,22,23,24 Besides, an activated antiapoptotic pathway contributes to tumor growth and progression.²⁵ Tumor hypoxia and accelerated angiogenesis have both been used as markers of poor prognosis for a variety of tumors, including breast cancer.^{10,11,12,13,22,26} Nonetheless, the role of autophagy remains, by and large, obscure in this respect. Is it a favorable or an unfavorable prognostic indicator in human breast malignancies?This study was designed to investigate this question in a series of 102 operable breast carcinomas. However, rather than using electron or fluorescence microscopy, this study uses light microscopy after immunohistochemical staining with an antibody recognizing the autophagy-related protein Atg8 (microtubule associated protein 1 light chain 3 MAP1LC3A or LC3A), an essential component of the autophagic machinery.^27,28 LC3A-II is derived from a proLC3 30-KDa protein after cleavage by autophagin Atg4 to produce the active cytosolic form LC3A-I (18 kDa). After activation by Atg7, LC3A-I is transferred to Atg3 and subsequently converted into the membrane-bound form LC3A-II.^8,28 The latter localizes on the isolation membranes and the complete spherical autophagosomal and autolysosomal membranes, forming a suitable marker of autophagic activity.     

Atrial fibrillation is associated with cardiac hypoxia

BackgroundAtrial fibrillation (AF), the most common human arrhythmia, is responsible for substantial morbidity and mortality and may be promoted by selective atrial ischemia and atrial fibrosis. Consequently, we investigated markers for hypoxia and angiogenesis in AF.MethodsRight atrial appendages (n=158) were grouped according to heart rhythm [sinus rhythm (SR) or AF]. The degree of fibrosis and microvessel density of all patients were determined morphometrically using Sirius-Red- and CD34/CD105-stained sections, respectively. Next, sections (n=77) underwent immunostaining to detect hypoxia- and angiogenesis-related proteins [hypoxia-inducible factor (HIF)1α, HIF2α, vascular endothelial growth factor (VEGF), VEGF receptor 2 (KDR), phosphorylated KDR (pKDR), carboanhydrase IX, platelet-derived growth factor] and the apoptosis-related B-cell lymphoma 2 protein.ResultsFibrosis progressed significantly from 14.7±0.8% (SR) to 22.3±1.4% (AF). While the positive cytoplasmic staining of HIF1α, HIF2α, VEGF, KDR, and pKDR rose significantly from SR to AF, their nuclear fractions fell (only pKDR significantly). The median CD34/CD105-positive microvessel size increased significantly from SR to AF.ConclusionsAF is closely associated with an atrial up-regulation of hypoxic and angiogenic markers. Whether this is cause, effect, or co-phenomenon of fibrosis remains to be investigated. It is conceivable that fibrosis might lead to an increased O₂ diffusion distance and thus induce ischemic signaling, which, in turn, leads to angiogenesis.     

Prognostic significance of microvessel density and other variables in Japanese and British patients with primary invasive breast cancer

The purpose of this study is to investigate the associations of microvessel density (MVD) and other pathological variables with survival, and whether they accounted for survival differences between Japanese and British patients. One hundred seventy-three Japanese and 184 British patients were included in the study. British patients were significantly older (56.3+/-11.4 years vs 52.5+/-12.9 years; P<0.01) and had smaller tumours (2.2+/-1.3 vs 2.7+/-1.8 cm; P<0.01), which were more frequently oestrogen receptor positive (78.8 vs 57.2%, P<0.01), had more grade III tumours (29.9 vs 21.4%, P=0.04) and more infiltrating lobular carcinomas (13.6 vs 4.0%, P<0.01) and a higher MVD compared with Japanese patients (57.9+/-19.8 vs 53.2+/-18.6; P=0.01). However, no difference in the prevalence of lymph-node metastasis was found between them (39.1 vs 37.5%, P=0.75). Younger British patients (age <50 years) had the highest MVD compared with Japanese and older British patients (P<0.01). Japanese patients were proportionately more likely to receive chemotherapy than endocrine therapy (P<0.01). British patients had a significantly worse relapse-free survival and overall survival compared with Japanese patients, after statistical adjustment for variables (hazard ratio=2.1, 2.4, P<0.01, P<0.01, respectively), especially, in T2 stage, low MVD and older subgroup (HR: 3.6, 5.0; 3.1, 3.3; 3.2, 3.9, respectively), but only in ER negative cases (P=0.04, P=0.01, respectively). The present study shows that MVD contributes to the Japanese-British disparity in breast cancer. However, the MVD variability did not explain the survival differences between Japanese and British patients.     

The american college of rheumatology 1990 criteria for the classification of fibromyalgia

To develop criteria for the classification of fibromyalgia, we studied 558 consecutive patients: 293 patients with fibromyalgia and 265 control patients. Interviews and examinations were performed by trained, blinded assessors. Control patients for the group with primary fibromyalgia were matched for age and sex, and limited to patients with disorders that could be confused with primary fibromyalgia. Control patients for the group with secondary-concomitant fibromyalgia were matched for age, sex, and concomitant rheumatic disorders. Widespread pain (axial plus upper and lower segment plus left- and right-sided pain) was found in 97.6% of all patients with fibromyalgia and in 69.1% of all control patients. The combination of widespread pain and mild or greater tenderness in ⩾ 11 of 18 tender point sites yielded a sensitivity of 88.4% and a specificity of 81.1%. Primary fibromyalgia patients and secondary-concomitant fibromyalgia patients did not differ statistically in any major study variable, and the criteria performed equally well in patients with and those without concomitant rheumatic conditions. The newly proposed criteria for the classification of fibromyalgia are 1) widespread pain in combination with 2) tenderness at 11 or more of the 18 specific tender point sites. No exclusions are made for the presence of concomitant radiographic or laboratory abnormalities. At the diagnostic or classification level, the distinction between primary fibromyalgia and secondary-concomitant fibromyalgia (as defined in the text) is abandoned.