Cancer associated fibroblasts (CAFs) are important components of the tumor microenvironment. Through secreting of multiple growth factors, cytokines and proteases, CAFs play a significant role in regulating the recruitment and function of various innate immune cells and adaptive immune cells in tumor microenvironment. In addition, extracellular matrix secreted by CAFs can also promote the formation of immunosuppression and hypoxia of tumor microenvironment. Here, we review the progress on CAFs in regulation of immune cells and tumor immunity.
Cancer-Associated Fibroblasts ; Extracellular Matrix ; immunology ; Humans ; Neoplasms ; immunology ; physiopathology ; Tumor Microenvironment ; immunology

CD146 Antigen ; immunology ; Fibroblasts ; immunology ; pathology ; Humans ; Scleroderma, Systemic ; drug therapy ; immunology ; pathology

We trapped a rat (Rattus norvegicus) infected with Capillaria hepatica. At necropsy, grossly yellowish-white nodules (2-3 mm in diameter) were noted to be scattered on the liver's surface. Microscopically, granulomatous and fibrotic nodules that contained the eggs and/or adult worms of Capillaria hepatica were detected in the liver. Septal fibrosis was diffusely formed throughout the liver. There were a number of ED1-positive macrophages located in the sinusoids of the pseudolobules. On the double staining, myofibroblasts and mast cells were generally observed within the fibrous septa with the mast cells in close proximity to the myofibroblasts. We suggest that the interactions between macrophages, myofibroblasts and mast cells play a role in the septal fibrosis observed in rats infected by Capillaria hepatica.
Animals ; *Capillaria ; Enoplida Infections/immunology/parasitology/*veterinary ; Fibroblasts/immunology ; Liver/parasitology/pathology ; Macrophages/immunology ; Mast Cells/immunology ; Rats ; Rodent Diseases/*immunology/*parasitology/pathology

OBJECTIVETo study the effect of the cultured supernatant of human silicotic alveolar macrophages (AM) on the expression of the collagen type I in human embryonic lung fibroblasts.

METHODSHuman alveolar macrophages were collected from a silicotic patient by bronchoalveolar lavage and exposed to silicon dioxide for 18 h. Then the cultured supernatant were used to culture human embryonic lung fibroblasts for 6 h, 12 h, 18 h, 24 h, 36 h, 48 h, 72 h. Then detected collagen anabolism and secretion with (3)H-proline detected the expression of the procollagen type I in the fibroblast with immunological method detected the quantity of collagen Type I in FB supernatant with Western blot.

RESULTSThe anabolism and secretion of collagen were increased in cultured supernatant of silicotic AM exposed to SiO(2), Along with the time, the expression of collagen type I increased. In cultured supernatant of silicotic AM exposed to SiO(2), ((3)H-proline: 1096.500 +/- 76.400, 707.000 +/- 62.160, OD: 0.314 +/- 0.011, OD: 14.218 +/- 0.342.

CONCLUSIONSiO(2) may affect the expression of collagen through AM mediation and participate in the formation of lung fibrosis.

Adult ; Cells, Cultured ; Collagen Type I ; metabolism ; Fibroblasts ; metabolism ; Humans ; Lung ; metabolism ; Macrophages, Alveolar ; immunology ; Male ; Silicosis ; immunology

Tumor microenvironment is defined as a heterogeneous complex composed of cancer cells, vascular endothelial cells, fibroblasts, and diverse immune cells. Cancer immunology is the study of interactions between the immune system and cancer cells which is applied to develop therapeutic strategies for human cancers. This review focused on tumor promoting myeloid derived cells such as tumor associated macrophages (TAM) and myeloid derived suppressor cells (MDSC) and their therapeutic applications.
Allergy and Immunology ; Endothelial Cells ; Fibroblasts ; Humans ; Immune System ; Macrophages ; Tumor Microenvironment

To study the correlation between 50% tissue-culture infective dose (TCID50) value and p27 antigen S/P value of Avian leukosis virus subgroup J and discuss their significance, chicken embryo fibroblast (CEF) cells were inoculated with Avian leukosis virus subgroup J strain NX0101 and samples were tested continuously for ten days after changing maintenance media. The correlation between TCID50 and p27 antigen S/P value of ten days were then analysized. Simultaneously, DF-1 cells were inoculated with NX0101 and passaged to 20 generations. Samples taken from 1st generation, 5th generation, 10th generation, 15th generation and 20th generation were tested for the TCID50 titer and the p27 antigen S/P value separately. A significant Pearson correlation was found between them in CEF cells (r = 0.85277; P < 0.0001) and in DF-1 cells (r = 0.93000; P = 0.0220). This study provided an important parameter for predicting TCID50 by detecting the p27 antigen S/P value.
Animals ; Avian Leukosis ; virology ; Avian Leukosis Virus ; immunology ; pathogenicity ; Chick Embryo ; Fibroblasts ; virology ; Proliferating Cell Nuclear Antigen ; analysis ; immunology ; Viral Load ; immunology

OBJECTIVETo investigate the effect of HCMV infection on the expression of interleukin-8 (IL-8) and regulated on activation, normal T expressed and secreted (RANTES) so as to explore the possible mechanism of the immune-pathological changes caused by HCMV infection.

METHODSExpression of both IL-8 and RANTES mRNA was detected by RT-PCR. Secretion of IL-8 and RANTES protein was quantitated by enzyme linked immune-sororbant assay (ELISA).

RESULTSAfter HCMV infection, the expression of IL-8 in HEL cells was found to be increased gradually concomitant with the prolonged infection time at both the transcriptional level and the protein level. By the time of 72 h.p.i, as compared with mock-infected cells, IL-8 mRNA expression was increased up to 12-fold and IL-8 protein up to 9-fold in HCMV-infected cells. The expression of RANTES mRNA was occurred at 8 h.p.i, with a maximum at 24 h.p.i, since then it remained relatively high level. The expression of RANTES protein peaked at 24 h.p.i, then dropped sharply and was almost undetectable by the time at 72 h.p.i.

CONCLUSIONHCMV infection of HEL cells may induce the transcription of IL-8 gene as well as the production of IL-8 orotein. HCMV may down-regulate extra-cellular production of RANTES protein.

Cells, Cultured ; Chemokines ; genetics ; immunology ; Cytomegalovirus ; physiology ; Cytomegalovirus Infections ; genetics ; immunology ; virology ; Female ; Fibroblasts ; immunology ; virology ; Gene Expression ; Humans ; Pilot Projects ; Pregnancy

Rheumatoid arthritis (RA) and osteoarthritis (OA), two common types of arthritis, affect the joints mainly by targeting the synovium and cartilage. Increasing evidence indicates that a significant network connects synovitis and cartilage destruction during the progression of arthritis. We recently demonstrated that hypoxia-inducible factor (HIF)-2alpha causes RA and OA by regulating the expression of catabolic factors in fibroblast-like synoviocytes (FLS) or chondrocytes. To address the reciprocal influences of HIF-2alpha on FLS and chondrocytes, we applied an in vitro co-culture system using a transwell apparatus. When co-cultured with HIF-2alpha-overexpressing chondrocytes, FLS exhibited increased expression of matrix metalloproteinases and inflammatory mediators, similar to the effects induced by tumor-necrosis factor (TNF)-alpha treatment of FLS. Moreover, chondrocytes co-cultured with HIF-2alpha-overexpressing FLS exhibited upregulation of Mmp3 and Mmp13, which is similar to the effects induced by interleukin (IL)-6 treatment of chondrocytes. We confirmed these differential HIF-2alpha-induced effects via distinct secretory mediators using Il6-knockout cells and a TNF-alpha-blocking antibody. The FLS-co-culture-induced gene expression changes in chondrocytes were significantly abrogated by IL-6 deficiency, whereas TNF-alpha neutralization blocked the alterations in gene expression associated with co-culture of FLS with chondrocytes. Our results further suggested that the observed changes might reflect the HIF-2alpha-induced upregulation of specific receptors for TNF-alpha (in FLS) and IL-6 (in chondrocytes). This study broadens our understanding of the possible regulatory mechanisms underlying the crosstalk between the synovium and cartilage in the presence of HIF-2alpha, and may suggest potential new anti-arthritis therapies.
Animals ; Arthritis/genetics/*immunology/pathology ; Arthritis, Rheumatoid/genetics/immunology/pathology ; Basic Helix-Loop-Helix Transcription Factors/genetics/*immunology ; Cells, Cultured ; Chondrocytes/immunology/metabolism/*pathology ; Coculture Techniques ; Fibroblasts/immunology/metabolism/*pathology ; Gene Expression Regulation ; Interleukin-6/genetics/*immunology ; Male ; Mice ; Mice, Inbred C57BL ; Osteoarthritis/genetics/immunology/pathology ; Synovial Membrane/immunology/metabolism/*pathology ; Tumor Necrosis Factor-alpha/genetics/*immunology ; Up-Regulation

To compare the anti-tumor effects of transmembrane TNF-alpha (TM-TNF) and secreted TNF-alpha (S-TNF) in vivo, mouse fibroblasts NIH3T3 were transfected separately with three types of retrovirus containing wild type TNF-alpha (Wt-TNF), TM-TNF mutant (TM-TNFm), S-TNF mutant (S-TNFm). Southern blot, RT-PCR, FACS and bioassay were used to investigate TNF-alpha gene integration, expression and its biological activity. It was found that both fixed cells and supernatant of NIH3T3/Wt-TNF, the fixed cells of NIH3T3/TM-TNFm and the supernatant of NIH3T3/S-TNFm could express high level of TNF-alpha or its mutants and effectively kill H22 in vitro. The transfected NIH3T3 were separately injected into the mice at the sites of H22 tumor cell inoculation according to a ratio of 5:1 or 1:1 (effector/target cells, E/T) after the third day of H22 challenge, respectively. At the E/T = 5:1, the NIH3T3/TM-TNFm induced the highest tumor regression, while NIH3T3/S-TNFm exerted the strongest tumor depressing effect at the E/T = 1:1 in vivo. No obvious side effects were noted throughout the course of treatment. The results suggest that both TM-TNF and S-TNF could cause tumor regression. The anti-tumor effect of TM-TNF would be more powerful and safe than that of S-TNF at the proper E/T ratio.
3T3 Cells ; Adenoviruses, Human ; genetics ; Animals ; Cytotoxicity, Immunologic ; immunology ; Fibroblasts ; cytology ; immunology ; Liver Neoplasms, Experimental ; immunology ; pathology ; Membrane Proteins ; secretion ; Mice ; Mutation ; Transfection ; Tumor Cells, Cultured ; Tumor Necrosis Factor-alpha ; genetics ; immunology ; secretion

Effect of interleukin-6 receptor (IL-6R) antibody on polymethyl methacrylate (PMMA) bone cement-mediated expression of osteoprotegerin (OPG) and receptor activator of nuclear factor-kappaB ligand (RANKL) in synovial fibroblasts was investigated. Synovial tissue obtained from total knee arthroplasty was digested and cultured. Inverted microscope was employed to observe the synovial cells and immunocytochemistry (SABC method) staining was used to identify synovial fibroblasts. This experiment was divided into three groups according to different culture media: PMMA group (75 μg/mL PMMA bone cement particles), IL-6R antibody group (10 ng/mL IL-6R antibody+75 μg/mL PMMA bone cement particles), and control group (no IL-6R antibody or PMMA bone cement particles). Influence of IL-6R antibody and PMMA on proliferation of synovial fibroblasts was measured by cell counting kit-8 (CCK-8). ELISA method was used to measure OPG and RANKL levels in culture solution. Fluorescence quantitative real-time PCR (FQ-PCR) was used to detect the expression of OPG and RANKL mRNA. After three consecutive passages, more than 95% of the primary synovial cells became long spindle fibroblast-like cells. SABC staining results showed that the fibroblast-like cells were negative for anti-CD68 antibody and positive for anti-vimentin antibody, with brown madder stained. CCK-8 test demonstrated that the absorbance (A) value at 450 nm was significantly lower in IL-6R antibody group than in PMMA group and control group (P<0.01), but there was no statistically significant difference in A value at 450 nm between the control group and PMMA group (P>0.05). Results of ELISA indicated that the expression of OPG was significantly higher in IL-6R antibody group than in PMMA group and control group (P<0.01). The expression of RANKL was inhibited (P<0.05), and the ratio of OPG/RANKL was significantly increased in IL-6R antibody group as compared with PMMA group and control group. There was no significant difference in the expression of OPG between control group and PMMA group (P>0.05), but the expression of RANKL was higher in PMMA group than in control group (P<0.05), and there was a significant difference in the ratio of OPG/RANKL between them (P<0.05). Results of FQ-PCR revealed the expression of RANKL mRNA was significantly inhibited (P<0.01) and the expression of OPG mRNA was significantly increased (P<0.01) in IL-6R antibody group as compared with PMMA group and control group. The expression of RANKL mRNA was higher in PMMA group than in control group (P<0.05), but the expression of OPG mRNA had no significant difference between them (P>0.05). IL-6R antibody could significantly increase the expression of OPG, but inhibit the expression of RANKL, which might provide a theoretical basis of molecular biology for the prevention and treatment of aseptic loosening of prosthesis.
Antibodies ; administration & dosage ; immunology ; Bone Cements ; Fibroblasts ; immunology ; Gene Expression ; drug effects ; Humans ; Osteoprotegerin ; biosynthesis ; genetics ; Polymethyl Methacrylate ; administration & dosage ; Prostheses and Implants ; RANK Ligand ; biosynthesis ; genetics ; metabolism ; Receptors, Interleukin-6 ; immunology ; metabolism ; Synovial Fluid ; immunology ; metabolism