Heterologous antisera against stromal mechanocytes of bone marrow

Heterologous rabbit antifibroblast serum (AFS) against stromal fibroblasts of guinea pig bone marrow was obtained. Both in the cytological reaction for adherent cells and in the indirect immunofluorescence test AFS specifically bound fibroblasts and their precursor cells (but not macrophages) in monolayer primary cultures of bone marrow, thymus, and spleen cells, and also precursor cells of fibroblasts of the blood and peritoneal exudate of guinea pigs (in the cytotoxic reaction). Fibroblast precursor cells of the thymus were more sensitive to the action of AFS than splenic and bone marrow cells, which suggests a higher concentration of common tissue-specific antigens on the stromal mechanocytes of the thymus. Precursor cells of stromal fibroblasts in native cell suspensions were essentially more sensitive to the cytotoxic action of AFS than colony-forming fibroblasts from passage cultures.Laboratory of Immunomorphology, N. F. Gamaleya Institute of Epidemiology and Microbiology, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR P. A. Vershilova.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 85, No. 4, pp. 451–454, April, 1978.     

Effect of IL-1 and TNF-α on growth of stromal fibroblasts in vitro

N. F. Gamaleya Research Institute of Epidemiology and Microbiology, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR S. V. Prozorovskii.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 113, No. 1, pp. 68–69, January, 1992.     

Effect of growth factors on stromal CFU-f colony formation in mouse bone marrow cell cultures

N. F. Gamaleya Research Institute of Epidemiology and Microbiology, Academy of Medical Sciences of the USSR, Moscow. Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 112, No. 12, pp. 615–617, December, 1991.     

Origin of stromal mechanocytes in bone marrow cultures

The strain of origin of fibroblasts growing in monolayer bone marrow cultures of semisyngeneic heterotopic grafts and radiochimeras was investigated by the indirect immunofluorescence method using an isoantiserum. All fibroblasts from bone marrow of complete radiochimeras were shown to be of recipient origin whereas histocyte—macrophages in the same cultures were of donor origin. All fibroblasts in bone marrow cultures of heterotopic grafts belonged to the original donor, whereas 80% of the histiocytes were of recipient origin. This proves directly that the stromal mechanocytes and hematopoietic cells, and also the macrophages, belonged to different cell lines.Laboratory of Immunomorphology, N. F. Gamaleya Institute of Epidemiology and Microbiology, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR O. V. Barovan.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 82, No. 10, pp. 1270–1271, October, 1976.     

Adhesion of hematopoietic cells and lymphocytes to stromal mechanocytes in vitro

Adhesion of guinea pig bone marrow and spleen cells to stromal mechanocytes of medullary, splenic, and thymic origin, to peritoneal exudate fibroblasts, and also to macrophages was studied in monolayer cultures. Primary cultures and subcultures of fibroblasts were able to bind hematopoietic cells and lymphocytes, and this property was independent of the origin of the mechanocytes. Hematopoietic cells were much less adherent to macrophages. Adhesion of cells to mechanocytes is determined by the number of adhesion sites present on the surface of the fibroblasts. There are significantly more of these sites for adhesion of myeloid cells on the surface of stromal mechanocytes than for adhesion of lymphocytes.Laboratory of Immunomorphology, N. F. Gamaleya Institute of Epidemiology and Microbiology, Academy of Medical Sciences of the USSR, Moscow. Department of Medical Biophysics, Institute of Chemical Physics, Academy of Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR V. A. Vershilov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 88, No. 12, pp. 703–704, December, 1979.     

Effect of stromal fibroblasts on antibody formation in nonadherent spleen cultures

Stromal fibroblasts from monolayer cultures of thymus, spleen, and bone marrow cells, when added to nonadherent suspension cultures of spleen cells have a significant influence on the accumulation of antibody-forming cells (AFC). Stromal fibroblasts of thymic origin stimulate-AFC formation, whereas those of bone-marrow origin inhibit antibody formation in these cultures. Stromal fibroblasts of splenic origin have the same effect on AFC formation as adherent cells. Stromal fibroblasts irradiated in a dose of 5000 R have the same action as unirradiated cells. The action of stromal fibroblasts on antibody formation in culture is manifested only if they adhere to the surface of the culture vessel.Laboratory of Immunomorphology, N. F. Gamaleya Institute of Epidemiology and Microbiology, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR P. A. Vershilova.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 85, No. 6, pp. 699–701, June, 1978.     

Suppression of collagen arthritis in rats by heterologous anti-idiotypic antisera against anticollagen antibodies

Affinity-purified rat anti-type II collagen antibodies were used to prepare anti-idiotypic antibodies in rabbits. It has been demonstrated that such anti-idiotypic antibodies are capable of binding to anti-type II collagen antibodies in vitro. Intravenous administration of heterologous anti-idiotypic antisera at the time of immunization with type II collagen resulted in a significant suppression of anti-type II collagen antibody formation and the development of arthritis, although delayed-type hypersensitivity skin test response to type II collagen was not affected. However, treatment of rats with heterologous anti-idiotypic antisera at Day 7 after immunization was ineffective in altering disease expression. On the other hand, treatment with heterologous anti-idiotypic antisera had no significant suppressive effect on the incidence or severity of adjuvant arthritis. These results indicate that the effect of heterologous anti-idiotypic antisera directed toward anti-type II collagen antibodies is disease specific and is restricted to collagen arthritis.     

Monoclonal antibodies against mouse splenic stromal cells

A panel of rat monoclonal antibodies directed against mouse splenic stromal cells were isolated. These monoclonal antibodies were Immunohistochemically divided into four groups which reacted with non-lymphoid cells of the murine spleen; (1) in the white pulp, (11) at the marginal zone, (111) in the red pulp, and (IV) on the endothelium of splenic blood vessels. These monoclonal antibodies were studied Immunohistochemically In lymphoid organs by means of light and electron microscopy. Monoclonal antibodies SS-4 (group I) reacted with fibroblastic reticulum cells that were distributed only in the white pulp of the spleen and In the follicular areas of lymph nodes. The SS-4 staining cell, In clustered splenic stromal cells, formed colonies which Included a small number of Thy-1 positive lymphocytes. Therefore, we concluded that SS4 staining stromal cells comprise the lymphoid cornpartment. In contrast, monoclonal antibodies SS-1, SS-3 and SS-5 (group II) reacted with dendritic shaped cells in the marginal zone of the spleen. Examination of splenic extra-medullary hematopolesis in mice rescued by bone marrow transplantation after lethal irradiation revealed that SS-3 and SS-5 reacted with dendritic shaped stromal cells in clonal nodules of engrafted marrow in the red pulp. SS-3 and SS-5 staining cells could not be observed in physiologic hematopoiesis of non-transplanted mice. It was suggested that SS3 and SS-5 staining stromal cells are Involved in primitive hematopoiesls. Monoclonal antibodies SS2, SS-6 and SS-7 (group 111) mainly reacted with dendritic cells and macro-phages in the red pulp. Monoclonal antibodies SS-8 and SS-9 (group IV) reacted with endothelial cells of blood vessels and sinuses. These findings of heterogeneity in mouse splenic stromal cells are further evidence that specific micro-envlronments are composed by speclalired stromal cells.     

Adipose stromal cells-secreted neuroprotective media against neuronal apoptosis

Transplantation of pluripotent adipose stem/stromal cells (ASC) alleviates tissue damage and improves functional deficits in both stroke and cardiovascular disease animal models. Recent studies indicate that the primary mechanism of ASC-induced repair may not be directly related to tissue regeneration through differentiation, but rather through paracrine mechanisms provided by secreted pro-survival and repair-inducing trophic factors. In this study, we have found that ASC-conditioned medium (ASC-CM) potently protected cerebellar granule neurons (CGN) from apoptosis induced by serum and potassium deprivation. Neural cell protection was mostly attributable to activated caspase-3 and Akt-mediated neuroprotective pathway signaling. Specific neutralization of neurotrophic factor activity demonstrated that serum and potassium deprivation-induced Akt-mediated neuroprotection and caspase-3-dependent apoptosis were mainly modulated by IGF-1. These data suggest that of the many neuroprotective factors secreted by ASC, IGF-1 is the major factor that mediates protection against serum and potassium deprivation-induced CGN apoptosis. This study establishes a mechanistic basis supporting the therapeutic application of ASC for neurological disorders, specifically through paracrine support provided by trophic factor secretion.     

Paracrine effects of uterine leucocytes on gene expression of human uterine stromal fibroblasts

The endometrium contains a distinct population of immune cellsthat undergo cyclic changes during the menstrual cycle and implantation.The majority of these leucocytes are uterine NK (uNK) cells,however how these cells interact with uterine stromal fibroblastsremains unclear. We therefore investigated the paracrine effectof medium conditioned by uterine decidual leucocytes (whichare enriched for uNK cells) on the gene expression profile ofendometrial stromal fibroblasts in vitro using a cDNA microarray.Our results, verified by real-time PCR, ELISA and FACS analysis,reveal that soluble factors from uterine leucocytes substantiallyalter endometrial stromal fibroblast gene expression. The largestgroup of up-regulated genes found was chemokines and cytokines.These include IL-8, CCL8 and CXCL1, which have also been shownto be stimulated by contact of stromal fibroblasts with trophoblast,suggesting that uNK cells work synergistically to support trophoblastmigration during implantation. The decidual leucocytes alsoup-regulated IL-15 and IL-15R in stromal fibroblasts which couldproduce a niche for uNK cells allowing proliferation withinand recruitment into the uterus, as seen in bone marrow. Overallthis study demonstrates, for the first time, the paracrine communicationbetween uterine leucocytes and uterine stromal fibroblasts,and adds to the understanding of how the uterine immune systemcontributes to the changes seen within the cycling endometrium.     

Antibody against rabbit immunoglobulin VH and L chain allotype determinants in heterologous anti-Ig antisera.

Heterologous anti-rabbit IgG antisera have been examined by radioimmunoassay for the occurrence of anti-VH antibody. Most of the sera contained antibody specific for VH allotype determinants (Aal). The antisera contained as much or more anti-al antibody as estimated in conventional alloantisera. It has further been shown that the anti-L chain antibody in all these sera was specific for L chain allotype determinants. Antibody against H chain-dependent L chain conformational determinants or cross-reacting determinants in CHlgamma and CHlalpha could not be demonstrated.     

Positive association between serum thymic stromal lymphopoietin and anti-citrullinated peptide antibodies in patients with rheumatoid arthritis

Thymic stromal lymphopoietin (TSLP) has been suggested recently to play an important role in the pathophysiology of rheumatoid arthritis (RA). However, there is little information on serum TSLP concentrations in RA and its clinical significance. The present study investigated whether serum TSLP concentrations were affected in patients with RA. Using an enzyme-linked immunosorbent assay (ELISA), we measured TSLP concentrations in the serum obtained from 100 patients with RA, 60 patients with osteoarthritis (OA) and 34 healthy volunteers. We also investigated the correlation between serum TSLP concentrations and clinical parameters of disease activity in RA [disease activity score using 28 joint counts (DAS28)-C-reactive protein (CRP), DAS28-erythrocyte sedimentation rate (ESR), Clinical Disease Activity Index (CDAI]), patient’s/-physician’s Visual Analogue Scale (VAS), swollen joints count, tender joints count, CRP, ESR and matrix metalloproteinase-3 (MMP-3) concentrations]. In addition, we investigated the correlation between serum TSLP concentrations and anti-citrullinated peptide antibody (ACPA) and serum tumour necrosis factor (TNF)-α. Serum TSLP levels in patients with RA were significantly higher than those in patients with OA and in healthy volunteers. Interestingly, serum TSLP concentrations were correlated significantly with ACPA titres, but not with other clinical parameters. There was a significant increase in serum TSLP concentrations in patients with RA, which was correlated positively with serum ACPA titres. These findings suggest that in patients with RA, TSLP may play a role in ACPA production by B cells.     

Breast stromal fibroblasts from histologically normal surgical margins are pro‐carcinogenic

There is evidence that normal breast stromal fibroblasts (NBFs) suppress tumour growth, while cancer‐associated fibroblasts (CAFs) promote tumourigenesis through functional interactions with tumour cells. Little is known about the biology and the carcinogenic potential of stromal fibroblasts present in histologically normal surgical margins (TCFs). Therefore, we first undertook gene expression analysis on five CAF/TCF pairs from breast cancer patients and three NBF samples (derived from mammoplasties). This comparative analysis revealed variation in gene expression between these three categories of cells, with a TCF‐specific gene expression profile. This variability was higher in TCFs than in their paired CAFs and also NBFs. Cytokine arrays show that TCFs have a specific secretory cytokine profile. In addition, stromal fibroblasts from surgical margins expressed high levels of α‐SMA and SDF‐1 and exhibited higher migratory/invasiveness abilities. Indirect co‐culture showed that TCF cells enhance the proliferation of non‐cancerous mammary epithelial cells and the epithelial‐to‐mesenchymal transition of breast cancer cells. Moreover, TCF and CAF cells increased the level of PCNA, MMP‐2 and the phosphorylated/activated form of Akt in normal breast luminal fibroblasts in a paracrine manner. Furthermore, TCFs were able to promote the formation and growth of humanized orthotopic breast tumours in nude mice. Interestingly, these TCF phenotypes and the extent of their effects were intermediate between those of NBFs and CAFs. Together, these results indicate that stromal fibroblasts located in non‐cancerous tissues exhibit a tumour‐promoting phenotype, indicating that their presence post‐surgery may play important roles in cancer recurrence. © 2013 The Authors. Journal of Pathology published by John Wiley & Sons Ltd on behalf of Pathological Society of Great Britain and Ireland.     

New developments in endometrial stromal sarcoma

Endometrial stromal tumours have been recently reclassified in the WHO 2014 Classification due to the discovery of new genetic fusions. This has enabled the subdivision of previously described undifferentiated endometrial sarcomas into the molecularly-defined high grade endometrial stromal sarcoma (HG ESS) and undifferentiated uterine sarcoma (UUS). In this review, we discuss the discoveries behind the 2014 Classification and its rationale, and give practical tips for diagnosis of these neoplasms, as well as discussing the differential diagnoses that one may consider.