muramidase and Melanoma

Myeloid Sarcoma with monocytic differentiation is rare and quite likely is missed by surgical pathologists. However it is frequently misdiagnosed because of its non-specific imaging and histological pattern.. We report the case of a 64-year-old woman with gastric primary myeloid sarcoma with monocytic differentiatio. Upper endoscopy revealed a neoplastic growth at the junction of the lesser curvature and gastric antrum. Except for a slightly increased peripheral monocyte count, no abnormalities were found on hematological and bone-marrow examination. Gastroscopic biopsy showed poorly differentiated atypical large cells with visible nucleoli and nuclear fission. Immunohistochemistry showed positive CD34, CD4, CD43, and CD56 expression, and weakly positive lysozyme expression. Immune markers for poorly differentiated adenocarcinoma, malignant melanoma, and lymphohematopoietic-system tumors were negative. The final diagnosis was myeloid sarcoma with monocytic differentiation. Chemotherapy did not shrink the tumor, so, radical surgery was performed. Although the tumor morphology did not change postoperatively, the immunophenotype did. CD68 and lysozyme expression (tumor tissue markers) changed from negative and weakly positive to strongly positive, AE1/3 expression (epithelial marker) changed from negative to positive, and CD34, CD4, CD43, and CD56 expression (common in naive hematopoietic cell-derived tumors) was greatly attenuated. Exome sequencing revealed missense mutations in FLT3 and PTPRB, which are associated with myeloid sarcoma, and in TP53, CD44, CD19, LTK, NOTCH2, and CNTN2, which are associated with lymphohematopoietic tumors and poorly differentiated cancers.. We diagnosed myeloid sarcoma with monocytic differentiation after excluding poorly differentiated adenocarcinoma, common lymphohematopoietic-system tumors, epithelioid sarcoma, and malignant melanoma. We identified that the immunophenotypic of patient had alterations after chemotherapy, and FLT3 gene mutations. We hope that the above results will improve our understanding of this rare tumor.

Topics: Adenocarcinoma; Cell Differentiation; Exome Sequencing; Female; Hematologic Neoplasms; Humans; Melanoma; Melanoma, Cutaneous Malignant; Middle Aged; Muramidase; Sarcoma, Myeloid

Tumor-associated macrophages (TAMs) are often associated with a poor prognosis in cancer. To gain a better understanding of cellular recruitment and dynamics of TAM biology during cancer progression, we established a novel transgenic mouse model for in vivo imaging of luciferase-expressing macrophages.. In vivo imaging of LysM-LG mice showed luciferase activity was generated by macrophages. Clodronate liposome-mediated depletion of macrophages lowered overall bioluminescence while lipopolysaccharide injection increased macrophage bioluminescence in both the B16 and ID8 models. Tracking macrophages weekly in tumor-bearing animals after intraperitoneal (i.p.) or intraovarian (i.o.) injection resulted in distinct, dynamic patterns of macrophage activity. Animals with metastatic ovarian cancer after i.p. injection exhibited significantly higher peritoneal macrophage activity compared to animals after i.o. injection.. The LysM-LG model allows tracking of macrophage recruitment and activation during disease initiation and progression in a noninvasive manner. This model provides a tool to visualize and monitor the benefit of pharmacological interventions targeting macrophages in preclinical models.

Topics: Animals; Disease Models, Animal; Female; Genes, Reporter; Luminescent Measurements; Macrophage Activation; Macrophages; Melanoma; Mice, Transgenic; Molecular Imaging; Muramidase; Ovarian Neoplasms; Promoter Regions, Genetic

The antibody-mediated targeted delivery of cytokines, growth factors and immunomodulators offers great potential for the therapy of cancer and other serious conditions. Interferon-alpha has long been used in the clinic for the treatment of patients with certain malignancies or with viral disease. Promising anticancer activity has recently been reported for two fusion proteins consisting of immunoglobulins bearing the interferon-alpha polypeptide at the C-terminal end of the molecule. Here we describe the design, production and characterization of a novel immunocytokine, in which murine interferon-alpha2 was sequentially fused with the tumor-targeting antibody fragment scFv(F8), specific to the alternatively-spliced EDA domain of fibronectin. The resulting fusion protein (F8-IFNa) could be produced to homogeneity and was shown to retain both antigen binding activity and interferon-alpha activity. Biodistribution studies in tumor-bearing mice with radioiodinated protein preparations confirmed the ability of F8-IFNa to selectively localize at the tumor site. However, using two different murine models of cancer (F9 teratocarcinomas and Cloudman S91 melanomas in immunocompetent mice), we could not detect a striking superiority for the therapeutic performance of F8-IFNa as compared to KSF-IFNa, a fusion protein of irrelevant specificity in the mouse which was used as negative control. In the paper, we present hypotheses why the antibody-based pharmacodelivery of interferon-alpha fails to eradicate tumors, in contrast to the situation observed by our group for other immunocytokines, which benefit from a selective localization at the tumor site.

Topics: Animal Structures; Animals; Antineoplastic Agents; Blood Vessels; Cell Line, Tumor; Cell Movement; Cell Proliferation; Female; Fibronectins; Humans; Immunoconjugates; Interferon-alpha; Leukocytes, Mononuclear; Melanoma; Mice; Mice, 129 Strain; Muramidase; Neoplasms; Neovascularization, Pathologic; Recombinant Fusion Proteins; Single-Chain Antibodies; Teratocarcinoma; Tissue Distribution; Treatment Outcome

To study the morphologic features, immunophenotypes, differential diagnoses and prognosis of histiocytic sarcoma (HS).. The clinical and pathologic findings of 6 cases of HS were reviewed. Immunohistochemical assay (Elivision staining) was also performed. Follow-up information was available in 4 patients.. There were altogether 3 males and 3 females. The age of patients ranged from 12 to 81 years old (median = 54.6 years). The sites of involvement included lymph node (number = 2 cases) and skin or soft tissue (number = 4 cases). The tumor was composed of sheets of large epithelioid cells with abundant eosinophilic cytoplasm, oval to irregular nuclei, vesicular chromatin and large nucleoli. Binucleated form was not uncommon. Two of the cases showed increased pleomorphism with multinucleated tumor giant cell formation. Focal cytoplasmic with foamy appearance was identified in 3 cases. One case demonstrated foci of spindly sarcomatoid appearance. Hemophagocytosis was identified in 2 cases. Mitotic figures were readily identified. The tumor cells were often accompanied by various numbers of inflammatory cells. Immunohistochemical study showed that all cases were diffusely positive for leukocyte common antigen, CD4, CD68 and CD163. Four of the 5 cases studied also expressed lysozyme. Amongst the 4 patients with follow-up information available, 3 died of the disease at 6 to 11 months interval after diagnosis. One patient, whose lesion was localized at the skin and soft tissue, survived for 3 years, with no evidence of tumor recurrence.. Accurate diagnosis of the HS is based on the combination of morphologic examination and immunohistochemical assay. HS often presents with clinically advanced disease and pursues an aggressive clinical course, with a poor response to therapy. However, a subset of cases presenting with clinically localized lesion may carry a relatively favorable long-term outcome.

Topics: Adult; Aged; Aged, 80 and over; Antigens, CD; Antigens, Differentiation, Myelomonocytic; Carcinoma, Renal Cell; Child; Diagnosis, Differential; Female; Follow-Up Studies; Histiocytic Sarcoma; Humans; Lymphoma, Large B-Cell, Diffuse; Lymphoma, Large-Cell, Anaplastic; Male; Melanoma; Muramidase; Prognosis; Receptors, Cell Surface; Skin Neoplasms; Soft Tissue Neoplasms; Young Adult

We report a 26-year-old male with a 4 mm diameter, asymmetric, irregularly pigmented and bordered, brown maculopapular lesion on the right nipple present since childhood with enlargement of the lesion within the last 3 months. Dermoscopy revealed a global globular pattern with the presence of focally light brown globules and irregular black globules in its centre. In vivo reflectance confocal microscopy (RCM) revealed dense junctional and dermal melanocytic nests of different sizes and shapes that appeared as sharply demarcated round to oval reflective structures; cellular outlines of single melanocytes were not always detected. In the centre of the lesion within the upper dermis, irregularly shaped, homogeneously reflecting structures were observed. As a clear differentiation between clusters of melanophages and melanocytic nests could not be made with certainty, an excisional biopsy was performed to establish the diagnosis of compound nevus with features of congenital nevus. Therefore, to prove that dermoscopic globules correlated with melanophages, the correlation between dermoscopic RCM and histopathology was necessary.

Topics: Adult; Antigens, CD; Antigens, Differentiation, Myelomonocytic; Dermoscopy; Humans; Male; Melanocytes; Melanoma; Microscopy, Confocal; Muramidase; Nevus, Pigmented; Nipples; Phosphoglucomutase; Skin Neoplasms; Treatment Outcome

The aim of this study was to evaluate the effects of lysozyme on the tumorigenicity of B-16V melanoma cells.. After performing a series of molecular biology applications, including mRNA isolation, reverse transcriptase polymerase chain reaction, restriction digestions and ligations, recombinant pHM6 vector harboring mouse lysozyme gene (pHM6mLys) was constructed. B-16V melanoma cells were transfected with plasmid DNAs (pHM6 and pHM6mLys). Transfected cells (B-16VpHM6 and B-16VpHM6mLys) were selected in media containing geneticin. B-16V, B-16VpHM6, and B-16VpHM6mLys cells were then injected subcutaneously (s.c.) to the three groups of C57BL/6 inbred mice (30 mice/group). These mice were examined every 3 days for s.c. tumor development over 41 days. The results were evaluated by using statistical methods.. Tumor formation was observed in all mice injected with B-16V and B-16VpHM6 cells in the first 8-12 days. However, tumor didn't develop in 16 of 30 of the mice injected with B-16VpHMmLys cells. Tumor-free animals (16 mice) in this group were reinjected with B-16V cells, and 9 of them died during the first 10 days of observation. Tumor development was not observed in the remaining 7 mice over 60 days of the experimental period. Results were statistically significant (p values < or = 0.05).. These findings indicate that lysozyme expressed by B-16VpHMmLys cells may suppress the tumorigenicity of these cells and may help development of protective immunity against B-16V melanoma cells.

Topics: Animals; Cloning, Molecular; Gene Expression Regulation, Neoplastic; Genetic Therapy; Genetic Vectors; Gentamicins; Melanoma; Melanoma, Experimental; Mice; Models, Statistical; Muramidase; Odds Ratio; Plasmids; Time Factors; Treatment Outcome

Previous studies have shown that immunohistochemical stains for histiocytes are immunoreactive for melanomas. Accordingly, their value in differentiating histiocytes and histiocytic lesions from melanomas was questioned. PG-M1, the most specific histiocytic marker, was not evaluated in these studies. Our aims were to assess the reactivity of PG-M1 with a series of primary cutaneous and metastatic melanomas and to establish the potential usefulness of this antibody in the differentiation between histiocytes and histiocytic tumors and melanomas. PG-M1 staining was performed in 50 primary cutaneous and metastatic melanomas. For comparison, additional sections were stained with KP-1 and lysozyme (commonly used as histiocytic markers) and with S-100 and HMB-45 (commonly used as melanoma markers). The intensity (1+, 2+) and extent (1+ to 4+) were recorded semiquantitatively. PG-M1 stained weakly (1+) and focally (2+) only four cases of melanoma (8%). In contrast, histiocytes were strongly reactive for PG-M1 in all cases, being readily differentiated from melanoma cells including the positive cases. KP-1 stained melanoma cells in 44 cases (88%), lysozyme in 11 cases (22%), S-100 in 50 cases (100%), and HMB-45 in 48 cases (96%). No changes were found after restaining of selected KP-1 and lysozyme positive melanomas using an endogenous avidin/biotin blocking kit. PG-M1 is helpful in discriminating histiocytes and histiocytic lesions from melanoma cells. We recommend its inclusion in any antibody panel put together to distinguish between them.

Topics: Antigens, CD; Antigens, Differentiation, Myelomonocytic; Biomarkers, Tumor; Diagnosis, Differential; Histiocytes; Humans; Immunohistochemistry; Melanoma; Muramidase; Neoplasms, Fibrous Tissue; Skin Neoplasms

We have previously demonstrated that class II-transfected tumor cells are very effective immunogens that protect against wild-type primary and metastatic tumor and, if supertransfected with genes encoding co-stimulatory molecules, are immunotherapeutic agents that successfully treat mice with established solid tumor. These results are consistent with our hypothesis that the class II-transfected tumor cells act as antigen-presenting cells (APCs) that directly activate tumor-specific CD4+ T cells; however, direct data supporting this hypothesis are lacking. In the present study, we test this hypothesis using class II-transfected tumor cells supertransfected with the hen egg lysozyme gene as a surrogate tumor antigen. In vitro antigen presentation assays demonstrate that class II-transfected tumor cells present to CD4+ T cells endogenously encoded tumor antigen, provided they do not co-express the class II-associated invariant chain. In vivo experiments using genetically marked tumor cells and host APCs demonstrate that both class II-transfected tumor cells and host cells are APCs for tumor-encoded antigens, although tumor cells appear to dominate the response. These results support the hypothesis that the immunogenicity and therapeutic value of class II-transfected tumor cells stem from their ability to function as APCs for tumor-encoded antigens and directly activate tumor-specific CD4+ T lymphocytes.

Topics: Animals; Antigen-Presenting Cells; Antigens, Neoplasm; Breast Neoplasms; CD4-Positive T-Lymphocytes; Gene Expression; H-2 Antigens; Histocompatibility Antigens Class II; HLA-DR Antigens; Humans; Immunization; Melanoma; Mice; Mice, Inbred A; Mice, Inbred C57BL; Muramidase; Sarcoma, Experimental; Transfection; Tumor Cells, Cultured

ANLL followed a brief period of aplastic anemia in a man treated intensively 4 years and 3 months previously with dacarbazine as "adjuvant" therapy for malignant melanoma. This is the first reported instance in which the latency between drug exposure and onset of leukemia strongly implicates dacarbazine as a leukemogenic agent.

Topics: Acute Disease; Dacarbazine; Humans; Leukemia; Male; Melanoma; Middle Aged; Muramidase; Scalp; Skin Neoplasms

Serum lysozyme activity (SLA) was measured in a turbidimetric assay with a microcentrifugal analyzer. In a control group of 53 healthy dogs of both sexes and ranging in age from 4 to 10 years, SLA had a mean value of 1.2 mg/l with a range (+/- 2 SD) of 0.6 - 1.8 mg/l. In 80 dogs with a variety of neoplastic diseases the histopathological diagnosis was compared with the SLA value. SLA value was increased in 83% of the cases with malignant tumors and in 29% of the cases with benign tumors. Proper clinical examination is essential in differentiating between neoplastic disease and some interfering diseases, e.g. chronic dermatitis, pyometra and chronic nephritis. Measuring of SLA in dogs may be helpful in screening those animals with suspected malignancies.

Topics: Adenoma; Animals; Carcinoma; Dog Diseases; Dogs; Female; Immunodiffusion; Male; Mammary Glands, Animal; Melanoma; Muramidase; Neoplasms; Nephelometry and Turbidimetry; Reference Values; Sarcoma; Skin Neoplasms; Soft Tissue Neoplasms

Twelve cases of malignant spindle-cell and sarcomatoid tumours of the skin of debatable nature were studied by immunocytochemical methods, using four antisera which might help contribute to resolution of the problems. The initial diagnosis made on structural grounds was confirmed by immunocytochemistry in six of eight cases in which a specific diagnosis had been made (one melanoma, three squamous carcinomas and two atypical fibroxanthomas). One case, initially regarded as AFX was reclassified as a squamous carcinoma, while a further case of possible AFX could not be confirmed by immunocytochemical study. Of the four cases in which structural examination was inconclusive, two were identified as squamous carcinomas and one as a melanoma by virtue of tumour markers. The fourth case was an intriguing actin-rich tumour of uncertain nature. Immunocytochemistry, despite certain limitations, has a valuable role to play in the analysis of the problematic spindle-cell malignant and pseudomalignant tumours of the skin.

Topics: Actins; Aged; Carcinoma, Squamous Cell; Female; Fibroma; Humans; Immunoenzyme Techniques; Keratins; Male; Melanoma; Middle Aged; Muramidase; S100 Proteins; Skin; Skin Neoplasms

The mononuclear phagocyte is well established as an in vitro cytotoxic effector cell for certain human tumors. The mechanism(s) for this action remains unclear. Increased levels of lysozyme, a cationic enzyme synthesized in large amounts by mononuclear phagocytes, are associated with increased resistance to transplantable animal tumors. In this study, we provide evidence that human lysozyme, isolated from the urine of leukemic patients, has marked potentiating effects on human monocyte-tumor-cell cytocidal activity. In addition, lysozyme-exposed monocytes incorporate increased quantities of leucine, suggesting that monocytes are capable of amplifying their own metabolic activation by secreting an endogenous constituent. Tri-N-acetyl-glucosamine, a competitive inhibitor for the active site of lysozyme, inhibits cytocidal activity. Conversely, protamine, an extraneous albeit similarly positively charged molecule, increases monocyte-mediated tumor cytotoxicity; this protamine effect is negated by heparin. We conclude that lysozyme, at least partially by its positive charge, is capable of enhancing in vitro monocyte tumor cell cytotoxicity; its in vivo secretion may potentiate monocyte-tumor-cell interaction.

Topics: Acetylglucosamine; Antineoplastic Agents; Cytotoxicity, Immunologic; Dose-Response Relationship, Immunologic; Hexosephosphates; Humans; Leucine; Melanoma; Monocytes; Muramidase; Urinary Bladder Neoplasms

Topics: Animals; Bromodeoxyuridine; Cell Line; Cricetinae; DNA, Neoplasm; Histones; Kinetics; Melanoma; Mesocricetus; Muramidase; Peptides; Protamines; Protein Binding; Structure-Activity Relationship

Topics: Adult; Aged; BCG Vaccine; Female; Humans; Male; Melanoma; Middle Aged; Muramidase; T-Lymphocytes; Time Factors

Serum lysozyme has been demonstrated to be an indicator for macrophage activity in the tumor-bearing host. Therefore, we investigated lysozyme levels in the sera of 336 untreated tumor patients (121 malignant melanoma, 61 lung cancers, 70 cervical cancers, 49 breast cancers and 35 benign breast tumors, and 36 healthy controls). Patients with malignant melanoma and lung cancer had significantly higher lysozyme levels than the healthy controls. Within the clinical stages in melanoma, there was a decrease of lysozyme in stages II and III in comparison to stage I, but still above that of the control values. Patients with benign breast tumors had normal levels, whereas in breast cancer patients of stages I and II there was a significant reduction in the lysozyme levels. In stages III and IV no differences to the control group could be detected. In patients with cervical cancer (FIGO II and III) serum lysozyme levels were found to be within the normal range. From this study it can not be concluded that serum lysozyme reflects the immunological reactivity of the tumor bearer. Nevertheless, the reduced levels in stages I and II of breast cancer might point to an immunological defect.

Topics: Breast Neoplasms; Female; Humans; Lung Neoplasms; Melanoma; Muramidase; Neoplasms; Uterine Cervical Neoplasms

It has been reported that lymphocytes from cancer patients give positive responses to PPD, myelin basic protein, tumour basic protein, and certain histone fractions in the MEM test. The underlying mechanisms of the MEM test are poorly understood, but it is widely assumed that it detects immunological sensitization to specific antigenic determinants. The cross-reactivity experienced is interpreted as indicating shared antigenicity. Since all the stimulatory proteins are strongly basic we investigated an alternative explanation that responsiveness is a function of electrical charge by comparing the known stimulatory proteins in the MEM test with two others of similar basicity: lysozyme and cytochrome-C. We obtained highly significant stimulation with PPD, tryptophane peptide of myelin, and tumour basic protein using Mantoux + cancer patients, but found no response to other basic proteins. We failed to confirm the reported activity of histone F2a. Our results indicate that basicity alone is insufficient to elicit response, and strengthens the concept that the MEM test is measuring sensitization to the determinants shared by myelin and tumour basic protein.

Topics: Animals; Cytochrome c Group; Epitopes; Guinea Pigs; Histones; Humans; Immunologic Techniques; Lymphocytes; Macrophages; Melanoma; Muramidase; Neoplasm Proteins; Surface Properties; Tuberculin

Serum lysozyme activity was measured in groups of untreated patients with malignant melanoma, hyperneophroma and breast carcinoma. Significant elevation of serum levels of the enzyme was confined to patients with localized disease. In the presence of metastatic disease such elevation was not detected. The rise in serum lysozyme activity was not due to renal damage or any infective process and in the case of malignant melanoma was shown to be associated with infiltration of the tumour mass by macrophages. In vitro studies demonstrated that the macrophages resident in a tumour mass are responsible for relasing lysozyme in large amounts. It is proposed that the elevation of serum lysozyme in these cases may be an indicator of macrophage-mediated host resistance and that the measurement of macrophage products such as lysozyme in the extracellular fluid may under well defined conditions provide useful clinical information concerning host reactions.

Topics: Adenocarcinoma; Adolescent; Adult; Breast Neoplasms; Cells, Cultured; Humans; Kidney Neoplasms; Macrophages; Melanoma; Middle Aged; Muramidase; Neoplasm Metastasis