Biomedical subjects
J A Reed
Publications and source records attributed to J A Reed.
CD34-reactive fibrous papule of the nose.
In human skin, the CD34 antigen is expressed on endothelium, periadnexal cells, and a population of reticular dermal interstitial cells. CD34 expression is characteristic of dermatofibrosarcoma protuberans and several other neoplasms, but not of typical fibrous papules of the nose. We describe a 16-year-old white girl with a slowly growing papule on the nose. Histopathology showed a dermal tumor with a superficial component of branched, thin-walled blood vessels and a deeper component of benign-appearing, spindle-shaped cells. These cells uniformly and strongly expressed CD34, but not factor XIIIa or markers of melanocytic, neural, muscular, vascular, or histiocytic differentiation. We consider this lesion a CD34-reactive fibrous papule. This benign tumor must be clearly distinguished from dermatofibrosarcoma protuberans, which also is composed of bundles of CD34-reactive spindle-shaped cells in most cases but has locally aggressive behavior.
Expression of the mast cell growth factor interleukin-3 in melanocytic lesions correlates with an increased number of mast cells in the perilesional stroma: implications for melanoma progression.
The molecular events responsible for tumor progression in human cutaneous malignant melanoma remain unclear; however, critical to the process is the dysregulated proliferation of tumor cells and the development of new vascular channels which allow further growth and dissemination. Connective tissue mast cells (MC) have been implicated in tumor progression because they concentrate around tumors (including melanomas) prior to the formation of new blood vessels, and because they contain many chemical mediators, including basic fibroblast growth factor (bFGF), known to have mitogenic and angiogenic effects. Several MC chemotactic and mitogenic factors have been described including interleukin-3 (IL-3). In order to determine whether there is a differential expression of this MC chemotactic/mitogenic factor with tumor progression in vivo, we evaluated by immunohistochemistry 85 melanocytic lesions including primary invasive malignant melanoma (PIMM), melanoma in situ (MMIS), and ordinary intradermal benign melanocytic nevi (BMN) for expression of IL-3. Nucleic acid in situ hybridization also was used to evaluate the melanocytic lesions for IL-3-specific mRNA transcripts. Intracellular IL-3 protein was detected in 29/33 (88%) PIMM and 15/25 (60%) MMIS, but was not detected in any (0/27; 0%) BMN (p < 0.0001). IL-3-specific mRNA transcripts were present in 3/4 PIMM and 2/10 MMIS in which IL-3 protein was not identified, but were not detected in any BMN. IL-3 mRNA or protein was not detected in normal melanocytes present in the perilesional epidermis of any of the specimens studied. Immunohistochemistry also was used to confirm the presence of IL-3 alpha-specific receptors on human cutaneous MC. As demonstrated by others, a significantly increased number of MC was present in the perilesional stroma of PIMM and MMIS vis a vis BMN (p < 0.0001). The results suggest that melanoma cells may attract MC in vivo by producing MC chemotactic/mitogenic factors such as IL-3. The recruitment of MC and the subsequent release of their potent mitogenic and angiogenic factors such as bFGF may thus represent a tumor-host interaction which favors tumor progression. Reed JA, McNutt NS, Bogdany JK, Albino AP. Expression of the mast cell growth factor interleukin-3 in melanocytic lesions correlates with an increased number of mast cells in the perilesional stroma: implications for melanoma progression.
Loss of expression of the p16/cyclin-dependent kinase inhibitor 2 tumor suppressor gene in melanocytic lesions correlates with invasive stage of tumor progression.
Sporadic and familial malignant melanoma susceptibility has been linked to defects in the chromosomal region 9p21. Recently, a putative 9p21 tumor suppressor gene, the cyclin dependent kinase inhibitor 2 (CDKN2) or p16 gene, has been shown to be deleted, mutated, or rearranged in a high percentage of sporadic melanoma cell lines, as well as mutated in the germline of a proportion of familial melanoma patients. CDKN2 encodes a M(r) 16,000 protein (p16) that plays a key role in cell cycle control by binding to the cyclin-dependent kinase 4 enzyme and inhibiting its ability to phosphorylate critical substrates necessary for transition past the G1 phase of the cell cycle. Thus, mutations or deletions of the CDKN2 gene could result in abnormal proliferation via defective cell cycle control. The correlation of 9p21 cytogenetic and molecular alterations with the clinical stages of melanoma progression suggests that dysfunction of a gene within this chromosomal region is critical to the evolution of melanoma. However, it remains unclear whether this gene is the CDKN2 gene. If so, then loss of p16 is potentially an initiating or early event in melanoma progression. To address the issues of what is the potential involvement of the CDKN2 gene in sporadic melanoma and precisely when during the clinically evident stages of melanoma progression defects in CDKN2 occur, we have evaluated by immunohistochemistry the expression of p16 protein in 103 melanocytic lesions representing all stages in the progression of melanoma. Our results suggest that loss of p16 protein expression is (a) not necessary for tumor initiation in malignant melanoma because all melanomas in situ and the majority of primary invasive melanomas retain expression of this protein; and (b) potentially more related to invasiveness or the ability to metastasize, because 52% of primary invasive tumors and 72% of metastatic lesions show partial or complete loss of expression of p16.
Differential expression of CD44 in malignant cutaneous epithelial neoplasms.
The CD44 antigen (ECMRIII, Hermes antigen) is a highly glycosylated cell-surface polypeptide involved in diverse cellular functions, including cell adhesion and lymphocyte-homing receptor activity. CD44 is also expressed in vivo by several tumors, including astrocytomas, meningiomas, and colonic adenocarcinomas. In addition, it has been shown that expression of CD44 appears to confer metastatic potential to cell lines derived from certain adenocarcinomas. In the skin, CD44 is normally expressed in epidermal keratinocytes and hair follicular, sebaceous, and eccrine epithelial cells. However, there have been few data with regard to the expression in vivo of CD44 in primary cutaneous neoplasms. Furthermore, there have not been studies in vivo of the possible differential expression of CD44 in primary versus metastatic tumors in the skin. We have examined by immunohistochemistry the expression of CD44 in cutaneous invasive and metastatic squamous cell carcinomas, metastatic adenocarcinomas, and basal cell carcinomas. All invasive and metastatic squamous cell carcinomas, as well as metastatic adenocarcinomas, strongly expressed CD44; however, basal cell carcinomas were nonreactive or showed only focal, minimal reactivity. Adjacent normal skin demonstrated CD44 immunoreactivity throughout the epidermis, including the basal layer. In addition, hair follicles and sebaceous and eccrine glands expressed CD44. The results suggest that expression of CD44 in these cutaneous epithelial tumors is not related to malignant transformation, but instead may be related to tumor progression and the ability to metastasize.
Immunohistochemistry of dermatofibromas and benign fibrous histiocytomas.
Dermatofibromas (DF) are common, benign skin tumors composed predominantly of cells having elongated nuclei and very scant cytoplasm (i.e., fibroblasts) and capillaries in a collagenous stroma. Some authors distinguish DF from benign fibrous histiocytomas (BFH), which are composed of cells with round to oval nuclei and abundant cytoplasm (i.e., histiocytes). In general, this group of tumors expresses factor XIIIa but not the antigen recognized by MAC 387. However, immunohistochemical differences specifically between DF and BFH have not been reported. We have studied the immunophenotype of 23 lesions having morphologic features predominantly either of DF (17 cases) or BFH (6 cases) using antibodies against desmin (muscle marker), alpha-smooth-muscle actin (muscle and myofibroblast marker), CD68 and HAM56 antigen (markers commonly expressed by macrophages, so called "histiocytic" markers), CD34 (a marker present in hematopoietic, vascular, and occasional dermal dendritic cells), and factor XIIIa (a transglutaminase present in many cells including dermal dendrocytes). Many spindle-shaped cells expressed alpha-smooth-muscle actin while many large, round cells expressed the histiocytic markers. However, most lesions expressed at least focally both alpha-smooth-muscle actin and "histiocytic" markers. Thus a clear-cut distinction between DF and BFH could not be made based on immunophenotype alone. Additionally, the prominent alpha-smooth-muscle actin immunoreactivity and desmin non-reactivity suggests myofibroblastic differentiation in the spindle-cell regions of these tumors, and indicates that expression of alpha-smooth-muscle actin cannot be used as definitive proof of muscle differentiation in spindle-cell tumors. We conclude that DF and BFH are not discrete entities, but represent polar expressions of one nosologic entity exhibiting both myofibroblastic and "histiocytic" differentiation.
Human cutaneous mast cells express basic fibroblast growth factor.
BACKGROUND: The normal response to tissue injury involves a complex sequence of inflammation and repair of epithelial and mesenchymal structures through a coordinated process of chemotaxis and cellular proliferation directed by specific chemical mediators. One mediator that is probably important to these processes is basic fibroblast growth factor (bFGF), a polypeptide present in diverse tissues and a potent chemotactic, mitogenic, and angiogenic factor in vitro. Although many cells have receptors for bFGF, the precise role of this factor as a paracrine mediator in vivo remains disputable because a mechanism for its secretion has not been elucidated. bFGF is known to be capable of binding to heparin, and our data demonstrate bFGF in mast cells (MCs) that also produce many other mediators (including heparin) important for wound healing and angiogenesis. EXPERIMENTAL DESIGN: We used nucleic acid in situ hybridization to detect bFGF-specific mRNA transcripts within MCs in formalin-fixed, paraffin-embedded specimens of nonlesional human skin and skin containing increased numbers of MCs, including granulation tissues, benign cutaneous mastocytosis, and solitary neurofibromas. Immunohistochemistry was used to confirm bFGF protein expression in each tissue studied. RESULTS: All tissues examined contained MCs that expressed bFGF mRNA transcripts. bFGF protein was also detected in MCs, but the intensity of labeling was dependent on prior treatment of the tissues with heparinase, suggesting that epitopes recognized by the bFGF-specific antiserum were masked by endogenous heparin. CONCLUSIONS: The results further elucidate the mechanism of involvement of MCs in tissue repair and angiogenesis and suggest that bFGF produced by MCs is bound to intracellular heparin. Thus, it seems possible that release of heparin by degranulation from MCs also may result in the release of biologically active bFGF in vivo.
Increased expression of basic fibroblast growth factor in squamous carcinogenesis of the head and neck is less prevalent following smoking cessation.
Basic fibroblast growth factor (bFGF), a potent angiogenic peptide, is thought to provide a growth advantage to a number of tumors including squamous cancer of the head and neck. The purpose of this study was to demonstrate the in situ expression of bFGF in lesions of the upper aerodigestive tract (UADT) and to correlate that with clinical parameters and known risk factors for carcinoma. On surgical specimens from 52 patients, we used a colorimetric in situ hybridization assay to determine the expression of bFGF mRNA in normal and pathologic conditions commonly seen in squamous mucosa. The extent of reactivity for the bFGF transcript was recorded on a subjective scale from 1+ to 3+, based on the visual intensity of labeling. These findings were subsequently correlated with clinical data. Basic fibroblast growth factor mRNA was detected at low to moderate levels in all sections of normal mucosa, with no distinction between patients with or without squamous cancer. Inflamed mucosa had comparatively strong expression of bFGF mRNA. Among lesions implicated in the stepwise nature of squamous carcinogenesis, we found increases in bFGF expression that were most significant at the level of carcinoma in situ, persisting through the invasive and nodal metastatic stages of the disease (P < 0.005). Interestingly, those increases were significantly less frequent among former smokers (P = 0.02). We have established the expression of bFGF mRNA in normal, inflammatory, and neoplastic tissue within the UADT. Furthermore, we note for the first time that increased expression is associated with the acquisition of more aggressive biologic behavior in squamous carcinogenesis.
Comparative changes in radial-bone density of elderly female lacto-ovovegetarians and omnivores.
A 5-y prospective study of the changes in radial-bone mineral density (BMD) of elderly white women (mean age, 81 y) living in four residential communities, including 49 Seventh-day Adventist lacto-ovovegetarians and 140 omnivores, was undertaken to determine the potential effects of usual dietary calcium in preventing the loss of BMD, measured by single-photon absorptiometry, at two radial sites. Changes in BMD and other variables from baseline (1983) to follow-up (1988) were: 1) mean calcium intakes in 1988 of 996 mg/d for omnivores and 733 mg/d for lacto-ovovegetarians changed little from 1983, 2) all women lost BMD (P < 0.05) over the 5 y period, 3) the annual BMD loss rates were approximately 1% at each site, 4) BMD loss was independent of calcium intake, 5) BMD loss rates were similar in both lacto-ovovegetarians and omnivores, and 6) the greater the loss of lean body mass, the greater the BMD loss (P < 0.05).
Morphologically variable bacilli of cat scratch disease are identified by immunocytochemical labeling with antibodies to Rochalimaea henselae.
The identification of the causative organisms of cat scratch disease (CSD) has been elusive. The demonstration of Warthin-Starry stain-positive pleomorphic bacilli in lymph nodes of patients with CSD and recent serologic and epidemiologic data suggest an etiologic role of Rochalimaea henselae in CSD. The authors studied lymph node biopsy specimens of 46 patients with illnesses clinically consistent with CSD and found pleomorphic bacilli in 15 (33%). The organisms were labeled by polyclonal rabbit antibodies induced by outer surface proteins of R henselae. This finding further supports the possibility of an important role of R henselae in the pathogenesis of CSD.
Primary cutaneous immunocytoma. A B-cell lymphoma that can easily be mistaken for cutaneous lymphoid hyperplasia.
It has been estimated that immunocytomas comprise roughly 2% of all cutaneous lymphomas. We studied five patients with primary cutaneous immunocytomas who presented with cutaneous nodules or plaques. Many of the infiltrates were "top-heavy" and polymorphous with admixed eosinophils, macrophages, lymphoid follicles, and non-neoplastic lymphocytes. Other potentially confusing findings were one case each of spongiotic dermatitis and leukocytoclastic vasculitis. The neoplastic cells were often situated at the peripheries of nodules and ranged from those with nuclei that resembled small lymphocytes to others that resembled immunoblasts. Most had eccentrically placed nuclei and fan-shaped cytoplasm. Monotypic kappa-light chain was found in all five cases, accompanied by gamma-heavy chain in two cases, and mu-heavy chain in one. In situ hybridization detected only kappa-mRNA in the four cases that yielded technically satisfactory results. The neoplastic cells did not express the B-cell antigen CD20; T-cells formed the centers of many nodules. Inappropriate staining for CD43 was evident in the neoplastic cells of one case. Because of reports of immunocytomas complicating acrodermatitis chronica atrophicans, we stained sections with an antiserum to Borrelia burgdorferi, which did not detect that organism. In situ hybridization did not detect EBER-1 RNA of the Epstein-Barr virus, which can be present in immunocytomas in immunocompromised patients. One patient died of disease after failing chemotherapy; another is alive with disseminated disease, and three are in remission following excision of lesions alone in two patients and chemotherapy in one patient who had relapsed following both excision and radiation therapy.
Immunohistochemical expression of BCL-2 in melanomas and intradermal nevi.
The BCL-2 gene is the prototype of a newly described family of oncogenes involved in tumorigenesis by blocking apoptosis, or programmed cell death. Overexpression of BCL-2 protein was originally described in follicular B-cell lymphomas bearing the 14;18 translocation. BCL-2 overexpression has also been described in other lymphomas and more rarely in neoplasms outside the lymphoid tissue. The aim of this paper is to determine the immunohistochemical expression of BCL-2 in intradermal nevi and primary invasive and metastatic melanoma. Formalin-fixed and paraffin-embedded tissues from 4 cutaneous melanoma metastases, 10 primary invasive melanomas, and 10 intradermal melanocytic nevi were immunolabeled with monoclonal antibodies directed against BCL-2 protein (Dako, clone 124) and Ki-67 antigen (Amac, clone MIB-1), after antigen retrieval techniques. Morphologically normal epidermal melanocytes expressed BCL-2, as did nevi and melanomas in virtually all cells. However, whereas the labeling in normal melanocytes and nevus cells showed a uniformly strong reactivity, melanoma cells showed a variable but mainly weak reactivity. Ki-67 antigen expression was restricted to melanomas. The widespread expression of BCL-2 suggests that this oncoprotein cannot be involved in the malignant transformation of melanocytic cells. It seems likely that the decreased BCL-2 expression detected in melanomas may reflect one further step of tumor progression in melanocytic neoplasms.
CD34 immunoreactivity distinguishes between scar tissue and residual tumor in re-excisional specimens of dermatofibrosarcoma protuberans.
Dermatofibrosarcoma protuberans (DFSP) is a locally aggressive tumor that often recurs after simple excision, and therefore usually requires wide surgical re-excision. It is sometimes difficult to distinguish histologically between residual tumor and scar tissue formed in response to the original biopsy. In an effort to solve this diagnostic problem, we have examined CD34 immunoreactivity in 10 primary biopsies of DFSP, 12 scars, and 7 re-excisional DFSP specimens containing residual tumor adjacent to scar tissue. In all 10 primary biopsies of DFSP, the tumor cells strongly and uniformly expressed CD34. In the 12 scars, only periadnexal cells, endothelial cells, and rare, scattered dendritic cells in the dermis were immunolabeled for CD34. In the 7 re-excisions of DFSP, the foci of residual DFSP were strongly immunolabeled, while the surrounding scar tissue was not. The pattern of CD34 immunoreactivity distinguishes DFSP from scar tissue, and thereby may permit more accurate assessment of surgical margins in re-excisional specimens of DFSP.
Rochalimaea henselae infection in acquired immunodeficiency syndrome causing inflammatory disease without angiomatosis or peliosis. Demonstration by immunocytochemistry and corroboration by DNA amplification.
Rochalimaea henselae causes bacillary angiomatosis, bacillary peliosis, and persistent bacteremia. Difficult to cultivate, it is detectable in infected tissues by immunocytochemistry. This technique demonstrated R henselae in autopsy specimens obtained from three deceased patients who had acquired immunodeficiency syndrome, with pathologic tissue changes lacking neoangiogenic features. From the first patient, the cause of nodular collections of lymphocytes and nonepithelioid histiocytes in the liver, spleen, lymph nodes, bone marrow, and heart eluded detection until immunocytochemical identification of R henselae. In the second case, unexpected gross and microscopic necroinflammatory nodules in the liver and spleen contained Warthin-Starry-staining bacilli identified as R henselae immunocytochemically. The third patient was found to have pathologic changes in his liver and spleen comparable with those of the second case, as well as several other disseminated infections. In two cases, identification of R henselae was corroborated through sequencing polymerase chain reaction-amplified bacterial DNA recovered from tissue; in one case, DNA could not be amplified, possibly because of postmortem degradation. Application of the immunocytochemical technique thus has expanded the recognized spectrum of histopathologic findings associated with R henselae infection in acquired immunodeficiency syndrome, as well as proving to be potentially more sensitive than DNA amplification for this purpose when applied to autopsy tissues.
Expression of transforming growth factor-beta 2 in malignant melanoma correlates with the depth of tumor invasion. Implications for tumor progression.
Transforming growth factor-beta 2 (TGF-beta 2) is a potent regulatory of proliferation and differentiation of both normal and malignant cells. In addition, TGF-beta 2 can exert a variety of immunosuppressive effects, suggesting that the production of this molecule contributes to impaired immunological surveillance of tumor development. In vitro, TGF-beta 2 expression has been demonstrated in cell lines derived from metastatic malignant melanocytes. but not in those derived from normal melanocytes. We sought to evaluate a potential role of TGF-beta 2 in the initiation or progression of malignant melanoma in vivo. We examined by nucleic acid in situ hybridization the expression of TGF-beta 2 mRNA transcripts in 124 melanocytic lesions including metastatic and primary invasive melanomas, melanomas in situ, nevi with architectural disorder and cytologic atypia, ordinary benign melanocytic nevi, and Spitz nevi. All metastatic melanomas and a majority (94%) of primary melanomas invasive to Clark's level III, IV, or V expressed TGF-beta 2 mRNA. A minority (41%) of Clark's level II primary invasive melanomas expressed this factor. All definitive melanomas in situ and nevi were negative. The results suggest that TGF-beta 2 expression in malignant melanoma may be a critical event in the development of deep invasion and metastases in malignant melanoma.
Differential expression of basic fibroblast growth factor (bFGF) in melanocytic lesions demonstrated by in situ hybridization. Implications for tumor progression.
Basic fibroblast growth factor (bFGF) is an angiogenic and mitogenic polypeptide produced by diverse cell types including cell lines derived from malignant melanomas but not from normal melanocytes. However, there is no consensus concerning in vivo expression of bFGF in melanocytic lesions due in part to the small numbers of cases studied to date. To evaluate further the possible differential expression of bFGF in melanocytic lesions, we examined 110 formalin-fixed, paraffin-embedded metastatic and primary invasive melanomas, melanomas in situ, nevi with architectural disorder and cytological atypia, and ordinary benign melanocyte nevi by nucleic acid in situ hybridization. All metastatic and primary invasive melanomas studied expressed bFGF mRNA, whereas melanomas in situ and benign melanocyte nevi were negative. Melanomas in situ with features of tumor regression and a majority of nevi with architectural disorder and cytological atypia also contained bFGF mrNA. The results suggest that in vivo bFGF expression is not requisite for malignant transformation per se, but appears to correlate more with invasion or fibroblastic reactions adjacent to the melanocyte lesions.
A 14/20 chromosome translocationp in Simmental cattle.
A 14/20 translocation was identified in 2 herds of Simmental cattle in the United States. All of the animals that carried the translocation had a common sire that was identified subsequently to carry the translocation. The semen of the bull had been used to inseminate a vast number of cows, so a substantial number of Simmental cattle in the United States may carry the 14/20 translocation. This translocation could have a detrimental effect on fertility in carrier animals; however, there are insufficient data about this particular abnormality to form conclusions. There appears to be sufficient evidence to urge caution in selecting carriers for breeding stock, especially when the potential exists for widespread dissemination of the abnormality through the use of artificial insemination.
Dual organism infection in biopsy specimens from immunocompromised patients: two cases demonstrated by immunocytochemistry.
We report two cases in which immunocytochemistry demonstrated simultaneous infections within a single biopsy specimen obtained from each of two immunocompromised patients in whom multiple organism infection was not suspected. Using a rapid immunocytochemical technique, Pneumocystis carinii and Cryptococcus neoformans were demonstrated in a transbronchial lung biopsy from the first patient; Candida and cytomegalovirus were demonstrated in an endoscopic biopsy of the esophagus from the second patient. Routine hematoxylin and eosin as well as special stains failed to establish the presence of more than one infectious agent in either case. Mycology cultures ultimately confirmed the presence of Cryptococcus in the first case, but cultures were not performed in the second case due to the scant amount of tissue obtained from the biopsy. The results indicate that concurrent infection by more than one opportunistic organism in immunocompromised patients can easily remain unrecognized. The use of immunocytochemistry may thus provide an important adjunct to routine and special stains or specific cultures.