T lymphocyte specificity of a lymphocyte-inhibiting factor (chalone) extracted from the thymus

The lymphocyte-inhibiting factor extracted from the thymus (LIFT) is effective in decreasing several immune responses. The aim of this study was to determine whether LIFT is able to depress humoral immune reactions and if so, whether it acts on T lymphocytes, B lymphocytes, or both. Simultaneous administration of LIFT and antigen severely impairs the IgG antibody formation against dinitrophenylated human IgG (DNP-HGG(a highly T lymphocyte-dependent response) and, to a lesser extent, the IgM antibody formation against sheep red blood cells (a partly T lymphocyte-dependent response). In contrast, the IgM anti-DNP-polymerized flagellin response (which does not require T lymphocyte collaboration) is not depressed at all by such treatment. These results demonstrate a LIFT action specificity on T lymphocytes only, and strongly argue for the assimilation of LIFT with a T lymphocyte chalone. The existence of a lymphocyte-inhibiting factor specific for B lymphocytes, or B lymphocyte chalone, is discussed.     

The effect of calf thymus extract (TFX) on human T lymphocyte and neutrophil mobility and chemotactic response in vitro

Calf thymus extract (TFX) was found to increase in vitro normal human T lymphocyte and granulocyte mobility, having no effect on their chemotactic response. TFX, when placed in the lower part of the chemotactic chamber was found to act as a chemotactic factor, even stronger than NHS and casein.     

The initial characterization of a thymus factor chemotactic to bone marrow cells

Thymus supernatants were produced by cluturing minced newborn CBA/J mouse thymuses in serum-free media for 48 h. Supernatants thus obtained were chemotactic to a subset of bone marrow cells as assessed in blind well chambers, and enriched for immature lymphoid cells in the migrating cell population. The enriched population of cells was shown to be capable of homing to the thymus of an irradiated mouse in vivo in a significantly higher percentage than nonmigrated bone marrow cells. In this report, initial characterization of the factor(s) responsible for this in vitro migration is presented. Several well studied thymic factors were compared with the thymus supernatants for their ability to induce migration of bone marrow cells in vitro. Thymulin (FTS-Zn), FTS, and TP-5 (the pentapeptide fragment of thymopoietin) were used. None of these factors demonstrated chemotactic properties in the migration assay using concentration ranges in which other in vitro activities have been observed. The chemoattractive activity of the supernatant was unaltered by ultracentrifugation. The effects of temperature on the chemotactic properties of thymus supernatant were examined, and a fifty percent decrease in observed migration occurred with thymus supernatant heated to 100 degrees C for 1 h. In addition, incubation of the supernatant for 1 h at 37 degrees C with chymotrypsin, but not with trypsin, inhibited migration, presumably by inactivation of the active factor. Using Amicon microconcentrators, the supernatant was separated into several fractions based on molecular weight. Initial data suggest that the active fraction is in the less than 10,000 mw range.     

Neutrophil chemotactic factor of anaphylaxis (NCF-A) release in aspirin-induced asthma

Neutrophil chemotactic activity (NCA) following oral challenge with aspirin (ASA) was determined in ASA-intolerant asthmatic subjects, and in ASA-tolerant asthmatic and normal subjects. There was a statistically significant fall in FEV₁ and a rise in NCA (P <0.01) following challenge in the ASA-sensitive subjects compared with that of the ASA-tolerant subjects and normal controls. No significant difference was observed between the latter two groups. The chemotactic factor identified in this study had a molecular weight greater than 150 000 which is consistent with NCF-A (neutrophil chemotactic factor of anaphylaxis). The ASA-induced fall in FEV₁ and rise in NCA was further studied in three of the ASA-intolerant asthmatic subjects, with and without pretreatment with inhaled sodium cromoglycate. In these subjects, the drug inhibited both the oral ASA-induced bronchoconstriction and the increase in neutrophil chemotactic activity. These results suggest that ASA-induced asthma involves mediator release from mast cells, as shown by the increase in NCA following ASA challenge and the protective effect of sodium cromoglycate which is considered to inhibit mast-cell degranulation.     

Antigen-induced eosinophil chemotactic factor (ECF) release by human leukocytes

A complement-independent eosinophil chemotactic factor (ECF) is described which is released from peripheral leukocytes of allergic and normal human volunteers after antigen stimulation and after exposure to anti-IgE. Dose response and time-release curves for ECF and histamine run closely parallel in this system. Histamine by itself is shown to have no effect on chemotaxis at the concentrations present in antigen-induced release, but is inhibitory at very high concentrations. Evidence suggests that the ECF released from human leukocytes is derived from basophils and is similar, or identical, to the ECF released from mast cells.This work was supported by Grants 07290 and 08270 from the National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland.Publication No. 197, O'Neill Research Laboratories.Dr. Czarnetzki is recipient of the Stetler Research Fund for Women Physicians.     

Lymphocyte-derived chemotactic factor synthesis in initial genital herpesvirus infection: correlation with lymphocyte transformation.

Lymphocyte transformation and production of lymphocyte-derived chemotactic factor in response to herpes simplex virus antigen were studied in 15 patients with initial genital herpes and 10 controls. The patients underwent frequent genital examinations, viral cultures, and weekly immunological studies for a period of 11 weeks. The production of lymphocyte-derived chemotactic factor was maximal in week 1 of the disease and declined to control levels by week 6. In contrast, lymphocyte transformation was lowest in week 1, reached a maximum by week 4, and declined to control levels by week 11. Production of lymphocyte-derived chemotactic factor in week 1 was significantly lower in nine patients who developed signs or symptoms of systemic herpes infection than in six who had localized disease. In addition, a marked but transient decline in the production of this lymphokine was observed in patients at the time of clinical recurrence. Virus-specific lymphocyte transformation correlated inversely with the duration of genital pain and lesions and did not correlate with the presence of systemic signs or symptoms. These findings indicate that during initial genital herpes infection the dynamics of lymphocyte transformation and those of lymphocyte-derived chemotactic factor production are different, and that the generation of this lymphokine is an early component of the cellular immune response in this disease. Furthermore, adequate produce of lymphocyte-derived chemotactic factor may be important in restricting herpes simplex virus to the genital area and preventing disease recurrence.     

雷藤舒(LLDT-8)对成纤维样滑膜细胞分泌趋化因子的影响

研究(5R)-5-羟基雷公藤内酯醇(雷藤舒,LLDT-8)对TNF-α联合IL-17诱导类风湿关节炎(rheumatoid arthritis,RA)成纤维样滑膜细胞(fibroblast-like synoviocytes,FLS)分泌趋化因子GRO-α、ENA-78、MCP-1、MIP-1α和RANTES的影响。ELISA法检测FLS上清液中GRO-α、ENA-78、MCP-1、MIP-1α和RANTES的浓度;RT-PCR法检测FLS中GRO-α、ENA-78、MCP-1、MIP-1α和RANTES mRNA的表达。结果显示LLDT-8可显著抑制TNF-α联合IL-17诱导的FLS上清液中GRO-α、ENA-78、MCP-1、MIP-1α和RANTES的表达;也可有效抑制TNF-α联合IL-17诱导的FLS GRO-α、ENA-78、MCP-1、MIP-1α和RANTES mRNA的表达。该研究提示LLDT-8通过抑制FLS产生GRO-α、ENA-78、MCP-1、MIP-1α和RANTES而在RA的治疗中发挥抗炎作用。     

Macrophage-released neutrophil chemotactic factor (MNCF) induces PMN-neutrophil migration through lectin-like activity

We have previously reported that rat peritoneal macrophages stimulated with LPS release a factor (MNCF) which induces neutrophil migration that is not blocked by glucocorticoids. The supernatant of macrophage monolayers stimulated with LPS was submitted to affinity chromatography on immobilized sugar columns. We observed that thed-gal binding fraction retained MNCF activity. This fraction, consisting of four protein components, was submitted to chromatography on Superdex 75, yielding a homogeneous preparation of the active component. MNCF has a MW of 54 KDa (gel filtration and SDS-PAGE) and pI<4.0 (isoelectrofocusing and chromatofocusing).d-gal did not interfere with the behaviour of known interleukins (IL-1β, IL-6, IL-8 TNF-α), but blocked MNCF activity in anin vitro migration assay. The present results reinforce our previous suggestion that MNCF may correspond to a novel monokine which induces neutrophil migration through a direct mechanism involving thed-gal binding site of the molecule.     

Macrophage-released neutrophil chemotactic factor (MNCF) induces PMN-neutrophil migration through lectin-like activity

We have previously reported that rat peritoneal macrophages stimulated with LPS release a factor (MNCF) which induces neutrophil migration that is not blocked by glucocorticoids. The supernatant of macrophage monolayers stimulated with LPS was submitted to affinity chromatography on immobilized sugar columns. We observed that thed-gal binding fraction retained MNCF activity. This fraction, consisting of four protein components, was submitted to chromatography on Superdex 75, yielding a homogeneous preparation of the active component. MNCF has a MW of 54 KDa (gel filtration and SDS-PAGE) and pI<4.0 (isoelectrofocusing=" and=">d-gal did not interfere with the behaviour of known interleukins (IL-1, IL-6, IL-8 TNF-), but blocked MNCF activity in anin vitro migration assay. The present results reinforce our previous suggestion that MNCF may correspond to a novel monokine which induces neutrophil migration through a direct mechanism involving thed-gal binding site of the molecule.     

CD28 signaling in neutrophil induces T-cell chemotactic factor(s) modulating T-cell response

We previously reported that human peripheral blood neutrophils express CD28 and interact with macrophage B7 to generate CD28 signaling through PI-3 kinase. Here, we demonstrate that crosslinking of CD28 on neutrophils results in the release of IFN-gamma, which restricts amastigote growth and modulates CD4+ T cells cytokine secretion. CD28 crosslinking also induces a T-cell chemotactic factor (TCF) that induces chemotactic migration of CD4+ T cells. Based on our previous and the current set of data, we propose an operational model explaining how neutrophils are involved in Leishmania infection and how the reported effect of neutrophils on the control of infection is mediated by alteration of T-cell function.     

Low molecular weight eosinophil chemotactic factor (ECF) in the serum of murine schistosomiasis japonica

Eosinophil chemotactic activity was detected in the serum obtained at an acute stage of murine schistosomiasis japonica. Gel filtration of the dialyzable fraction of the serum on Sephadex G25 showed that the chemotactic component had an apparent molecular weight of less than 1,000. It was stable to heating, but was sensitive to pronase or carboxypeptidase A digestions, indicating its peptide nature. Eosinophil chemotactic activity of the dialysate of the serum from mast cell-deficient W/Wv mice infected with Schistosoma japonicum was far less than that from normal littermate +/+ mice, although the titers of specific IgE antibody to soluble egg antigen in the serum measured by passive cutaneous anaphylaxis was comparable between them. These results suggest that at least some part of low molecular weight ECF in the circulation seems to be a ECF-A derived from mast cells. Possible biological significance of circulating ECF in schistosomiasis has been discussed.     

Effect of L-asparaginase and hydrocortisone on human lymphocyte transformation and production of a mononuclear leucocyte chemotactic factor in vitro

Human peripheral lymphocytes stimulated in vitro with PHA produce a soluble factor which is chemotactic for homologous monocytes. The synthesis of this factor was found to precede the blastogenic response as measured by [³H] thymidine incorporation. In cultures of unseparated leucocytes the maximum of chemotactic activity was detected after 24 hr, whereas in supernatants from purified lymphocyte suspensions the maximal synthesis occurs after 72 hr. High doses of L-asparaginase from E. coli which have been found to prevent lymphocyte transformation completely have no influence on the production of the chemotactic factor. Therefore it seems possible that the induction of DNA synthesis by PHA and its effect on the production of a chemotactic factor depend on different biochemical mechanisms. In contrast hydrocortisone leads to a dose-dependent inhibition of both DNA synthesis and chemotactic response.     

Generation of the eosinophil chemotactic factor (ECF) from various cell types by melittin

The peptide melittin, the main constituent of bee venom is a potent stimulus for the generation of an eosinophil chemotactic factor (ECF) from human polymorphonuclear neutrophils, rat mast cells and rat peritoneal cells depleted in mast cells. Optimal EFC induction required a sublytic activation of the cells. With each cell type the kinetics of ECF generation were similar in that after an early rise in activity a steep fall off occurred at later times of incubation suggesting a mechanism of inactivation. The induction of ECF by melittin is increased in the presence of calcium. The polar portion of the melittin molecule (aminoacids 20–26) is responsible for the generation of the chemotactic activity. Other peptides of honey bee venom such as the mast cell degranulating peptide (MCD) or apamine do not initiate ECF release. It appears that melittin leads to ECF induction via the phospholipase A₂-arachidonic acid dependent pathway of cell activation. Our data suggests that the lipid mediator ECF can be obtained from phagocytes and mast cells thus indicating the interdependence of inflammatory reactions.     

Normal chemotactic migration of polymorphonuclear leukocytes stimulated with mononuclear-derived chemotactic factor in ulcerative colitis

The migration of polymorphonuclear leukocytes preincubated with autologous or heterologous serum was examined in 100 patients with untreated ulcerative colitis (UC) and in 100 age- and sex-matched healthy controls. The activity of complement-derived chemotactic factors and mononuclear-derived chemotactic factor (MDCF) was also investigated in the same group of patients. No significant difference was found in random and chemotactic migration of patients or control leukocytes preincubated in different concentrations (10, 50 and 100%) of autologous or heterologous serum. Defective chemotaxis of leukocytes stimulated with complement-derived chemotactic factors was found in UC and was more marked in patients in remission than with active UC independently of whether complement was activated by the alternative or the classical pathway. However, the random migration of neutrophils was enhanced in both groups of UC patients. The leukocytes of patients stimulated with MDCF (mononuclear cells were activated with lipopolysaccharide from Escherichia coli O55:B5) show normal chemotaxis. Our data suggest an impairment of neutrophil receptors for complement-derived chemotactic factor in UC. The decreased response of neutrophils to these factors and normal response to MDCF suggest that the main way in which cells are attracted to the site of inflammation in UC may be a factor produced by stimulated mononuclear cells.