Macrophage Chemotactic Factor Research Articles

Reply: Adjuvant immunochemotherapy with oral Tegafur/Uracil plus PSK in patients with stage II or III colorectal cancer

Reply: Adjuvant immunochemotherapy with oral Tegafur/Uracil plus PSK in patients with stage II or III colorectal cancer

Cytokine regulation by peroxisome proliferator-activated receptor gamma in human endometrial cells

Objective To determine whether peroxisome proliferator-activated receptor (PPAR)-γ ligands can affect the expression of interleukin-6 (IL-6) and cytokines related to the pathogenesis of endometriosis. Design In vitro study to determine whether PPARs are expressed in human endometrial cells and determine the effects of various PPAR-γ ligands on IL-6 and other cytokine expression in these cells. Setting Academic medical center. Patient(s) Women presenting for infertility workup. Intervention(s) Endometrial cell cultures were treated with PPAR-γ ligands. Main outcome measure(s) Interleukin-6, IL-8, colony stimulating factor-1 (CSF-1) and macrophage chemotactic factor (MCP-1) protein secretion, messenger RNA expression of IL-6, PPAR-α, -β, and -γ. Result(s) Using a human endometrial cell line (EM42), as well as primary stromal and epithelial endometrial cells, we show the presence of PPAR-α, -β, and -γ by reverse transcription-polymerase chain reaction (RT-PCR) in these cells. PPAR-γ ligands stimulated IL-6 secretion and induced enhancement of IL-6 mRNA levels. These ligands also stimulated the secretion of IL-8 and CSF-1. Conclusion(s) PPAR-γ may play a role in the pathogenesis of endometriosis related to the production of IL-6 and some other cytokines.

Intraoperative transfection of vein grafts with the NFκB decoy in a canine aortocoronary bypass model: a strategy to attenuate intimal hyperplasia

Background The nuclear transcriptional factor NFκB is reported to play an important role in the expression of genes for neutrophil and macrophage chemotactic factors, adhesion molecules, and cell cycle–regulating proteins. In aortocoronary bypass surgery, the saphenous vein often develops vein graft disease. Here, we investigated whether transfection of a cis element decoy oligodeoxynucleotide of NFκB (NFκB decoy) into the vein graft wall suppresses neointimal hyperplasia and the differentiation of medial smooth muscle cells. Methods We established a canine aortocoronary bypass grafting model that has a saphenous vein graft between the left anterior descending coronary artery and the descending aorta. Pressure-mediated transfection of a scrambled (SD group; n = 5) or NFκB decoy (ND group; n = 5) into the graft wall was performed intraoperatively. The grafts were gently harvested at 4 weeks postoperative, and the middle portion of the graft was examined histopathologically. Results The average neointimal area of the ND group was significantly suppressed (SD group, 2.63 ± 1.00 mm 2 vs ND group, 0.88 ± 0.66, p < 0.05), and the differentiation and proliferation of the medial smooth muscle cells in the ND group were also suppressed (proliferating cell nuclear antigen index: SD group, 56 ± 24 vs ND, 13 ± 4, p < 0.05). Conclusions These results demonstrated the efficacy of intraoperative transfection of the NFκB decoy into the vein graft wall for attenuation of neointima formation.

Modulation of intracellular calcium and calmodulin by melatonin in MCF-7 human breast cancer cells.

The pineal hormone, melatonin, has been shown to inhibit the proliferation of the estrogen receptor alpha (ERalpha)-positive macrophage chemotactic factor (MCF)-7 human breast cancer cells. Previous studies from other systems indicate that melatonin modulates the calcium (Ca2+)/calmodulin (CaM) signaling pathway either by changing intracellular calcium concentration ([Ca2+]i) via activation of its G-protein coupled membrane receptors, or through a direct interaction with CaM. In this study, although melatonin alone had no effect on basal [Ca2+]i in MCF-7 cells, it significantly enhanced the elevation of [Ca2+]i induction by extracellular adenosine triphosphate (ATP), which increases [Ca2+]i via the G protein-coupled P2y-purinoceptor and the phospholipase C (PLC) pathway. Pretreatment of MCF-7 cells with 10(-7) M melatonin increased the 10(-5) M ATP-induced [Ca2+]i peak change from 79.4 +/- 11.6 nM to 146.2 +/- 22.3 nM. Furthermore, without changing total cellular CaM levels, melatonin markedly increased the amount of membrane-bound CaM to 237 and 162% of control levels after I and 6 hr of treatment, respectively. Cytosolic CaM levels were also elevated to 172% of control after 6 hr of melatonin treatment. Correlative growth studies demonstrated that ATP (10(-5) M) can stimulate MCF-7 cell growth, that melatonin can suppress MCF-7 cell proliferation, but that pretreatment of MCF-7 cells with melatonin followed by ATP(10(-5) M), like 10(-4) M ATP can further suppress MCF-7 cell proliferation; this indicates that melatonin's potentiation of ATP induced [Ca2+]i may be above the threshold for cell growth. Given the important role of [Ca2+]i and CaM in tumor cell homeostasis and proliferation and melatonin's modulation of [Ca2+]i, melatonin's effects on the Ca2+/CaM signaling pathway may play an important role in mediating the growth-inhibitory effect of melatonin on MCF-7 human breast cancer cells.

Antitumor effect of a peptide-glucan preparation extracted from Agaricus blazei in a double-grafted tumor system in mice.

The antitumor effect of extracts obtained from the fruit body of Agaricus blazei Murill was examined in a double-grafted tumor system, in which BALB/c mice received simultaneous intradermal injections of Meth-A tumor cells in both the right (10(6) cells) and left flank (2 x 10(5) cells), and were then injected with 5 mg of extracts of A. blazei in the right tumor on days 3, 4 and 5. Intratumoral administration of ethanol-soluble (Fraction 1), water-ethanol-soluble (Fraction 2), ammonium oxalate-soluble (Fraction 3) and ammonium oxalate-insoluble (Fraction 4) fractions resulted in inhibition of tumor growth, with Fraction 3 showing the most tumoricidal activity, producing regression of the right tumor and inhibition of growth of the left, non-injected tumor. The maximum effect was obtained using 0.5 mg of Fraction 3 and this amount was used in subsequent experiments. The antitumor effect of intratumorally administered Fraction 3 was enhanced by oral ad lib administration of feed containing 0.083% of Fraction 3. When immunized spleen cells from mice that had been cured by intratumoral administration of 0.5 mg of Fraction 3 were directly injected (2 x 10(7) cells/mouse) into the Meth-A tumor, tumor growth was inhibited. The tumor cells on day 7 from the Fraction 3-treated right tumor and from the left tumor were cultured for 24 h and their culture supernatants were assayed for neutrophil or macrophage chemotactic activity. Significant macrophage chemotactic factor activity was detected in the culture media from the left tumor tissue. Serum levels of immunosuppressive acidic protein (IAP), produced by activated macrophages and neutrophils, increased transiently soon after intradermal injection of 0.5 mg of Fraction 3. These results suggest that regression of the left non-injected tumor was due to an immune reaction, involving induction of cytotoxic cells in the spleen, and the release of chemotactic factors in the distant tumor.

TCR alpha beta+ CD4- CD8- T cells differentiate extrathymically in an lck-independent manner and participate in early response against Listeria monocytogenes infection through interferon-gamma production.

T-cell receptor (TCR) alpha beta+ CD4- CD8- (double-negative; DN) T cells appear in the peritoneal cavity at an early stage of intraperitoneal (i.p.) infection with the intracellular pathogen Listeria monocytogenes. In the present report, we analysed the developmental pathway and functions of the TCR alpha beta+ DN T cells using the L. monocytogenes infection system. The TCR alpha beta+ DN T cells appeared in the peritoneal cavity after L. monocytogenes i.p. infection in adult-thymectomized lethally irradiated bone marrow chimeras and p56lck-deficient mice. The results demonstrated that the TCR alpha beta+ DN T cells can develop extrathymically in a p56lck-independent manner. Reverse transcription-polymerase chain reaction (RT-PCR) analysis showed that the TCR alpha beta+ DN T cells expressed genes for interferon-gamma (IFN-gamma), the macrophage chemotactic factors MCP-1 and Eta-1, and granulocyte-macrophage colony-stimulating factor (GM-CSF) but lacked expression of genes for interleukin-2 (IL-2), IL-4 and IL-10. As expected from the RT-PCR analysis, the TCR alpha beta+ DN T cells produced IFN-gamma in response to anti-TCR beta monoclonal antibody (mAb), anti-CD3 mAb and L. monocytogenes-infected macrophages but IL-4 was undetectable after the stimulation. Furthermore, the intracellular cytokine staining analysis demonstrated that approximately half of the TCR alpha beta+ DN T cells detectable at the early stage of L. monocytogenes infection were IFN-gamma-producing cells. All of the results suggest that the TCR alpha beta+ DN T cells develop through a unique extrathymic p56lck-independent pathway and participate in early protection against bacterial infection through activation and accumulation of macrophages.

Relationship between induction of macrophage chemotactic factors and formation of granulomas caused by mycoloyl glycolipids from Rhodococcus ruber (Nocardia rubra).

Mycoloyl glycolipids cause granulomas in the lungs, liver, and spleen of mice, but the mechanism is not fully understood. To understand the role of macrophage chemotactic factors (MCFs) in granuloma formation, we prepared various mycoloyl glycolipids with different carbohydrate moieties: trehalose dimycolate (TDM), glucose mycolate (GM), mannose mycolate (MM), and fructose mycolate (FM) from Rhodococcus ruber, and examined the relationship between their MCF induction in peritoneal macrophages and the extent of granuloma formation. The molecular mass of each glycolipid was confirmed by fast-atom-bombardment mass-spectrometry. TDM or GM caused granulomas in the lungs, spleen, and liver of ICR mice, but MM and FM did not. The culture supernatant of peritoneal macrophages stimulated with TDM or GM increased macrophage migration, whereas MM and FM had no chemotactic activity. The activity of interleukin-1 (IL-1) in the supernatant was increased equally by each glycolipid and was therefore not related to chemotaxis. Tumor necrosis factor-alpha (TNF-alpha) and granulocyte-macrophage colony-stimulating factor (GM-CSF) were not detected in the four supernatants. The TDM-induced MCF was heat-stable, trypsin-labile, and undialyzable. Furthermore, we separated two MCF active fractions from the supernatant of TDM-stimulated macrophages by gel filtration. These factors acted on macrophages but not on neutrophils. Our results suggested that macrophages recognize the sugar moieties of mycoloyl glycolipids and may, in response, generate a MCF that may play an important role in the macrophage or monocyte recruitment which is essential prior to granuloma formation.

Influence of Surfactant on Cytokine Release from Ozone-Exposed Human and Bovine Alveolar Macrophages in Vitro

AbstractPrevious studies at our laboratory demonstrated that ozone, a ubiquitous air pollutant, was able to induce the liberation of cytokines from alveolar macrophages (AM) in vitro. As the lung cells in the alveoli are lined by surfactant, we exposed to ozone in this study surfactant-coated human (HAM) and bovine (BAM) alveolar macrophages in order to examine isolated cells under conditions that simulate as nearly as possible the in vivo situation in the lung. BAM were incubated with 0.25, 0.5, or 1 ppm ozone for 2 or 4 h in the presence or absence of the synthetic surfactant surrogate Alveofact or dipalmitoyl lecithine (DPL), the major lipid of natural surfactant. Incubations of BAM with Alveofact led, for 1 ppm ozone, to a significant suppression of the ozone-induced liberation of tumor necrosis factor α (TNF-α) and Mac-ChF, a chemotactic factor for macrophages, and DPL also reduced significantly the ozone-elicited Mac-ChF release when 0.5 ppm ozone was tested. When we exposed BAM to a combination of ...

Influence of Coexposure of Ozone with Quartz, Latex, Albumin, and LPS on TNF-α and Chemotactic Factor Release by Bovine Alveolar Macrophages in Vitro

Ambient air is a mixture of particles and gases, but in studies with air pollutants often a single compound is used. Thus, it is necessary to investigate the combined effect of pollutant mixtures. Bovine alveolar macrophages (BAM) were incubated in vitro for 2 h with quartz, latex, or albumin particles and afterward exposed to 0.25 or 1 ppm ozone. Incubations of BAM with 1 ppm ozone, quartz, albumin, or LPS alone significantly increased the release of TNF-α and a chemotactic factor for macrophages (Mac-ChF) from the cultured cells. Combinations of particles with ozone showed in no case a synergistic or antagonistic effect on the TNF-α or chemoattractant secretion. Incubations with LPS of ozone-pretreated cells resulted in an additive effect on Mac-ChF release. In conclusion, the results of this study provided evidence that the in vitro coexposure of ozone and particles does not result in an interaction, at least not for the release of TNF-α and Mac-ChF from BAM.

Phenotypic Properties and Cytokine Production of Skin-Infiltrating Cells Obtained from Guinea Pig Delayed-Type Hypersensitivity Reaction Sites

In this study, skin-infiltrating cells were prepared from guinea pig delayed-type hypersensitivity reaction sites, and their phenotypic properties and cytokine production were examined. Our cell preparations contained around 45% nonspecific esterase-positive cells and a few skin-derived cells. Skin-infiltrating cells produced TNF activity and macrophage chemotactic activity (MCA), whereas resident skin cells produced much lower activity. The highest TNF activity was produced by skin-infiltrating cells at 2 hr after antigen injection. In contrast, a larger amount of MCA was produced by skin-infiltrating cells at 24 and 48 hr than at 2 hr after antigen injection. These kinetics were in good agreement with our previous results regarding cytokine activity in skin extracts. Gel filtration analysis and neutralization assay with a monoclonal antibody against human macrophage chemotactic factor with a molecular weight of 1 kDa (MCF-1) indicated that a guinea pig homologue of human MCF-1 was partially responsible for the MCA.

Involvement of Inflammatory Cytokines in a Delayed-Type Hypersensitivity Reaction

We previously reported the amino acid sequence of the human macrophage chemotactic factor with a molecular weight of 1000 (MCF-1) was WLGREDGSE. In this study we examined the roles of MCF-1 and tumor necrosis factor (TNF) in a guinea pig delayed-type hypersensitivity (DTH) reaction. When macrophage chemotactic activity (MCA) in a skin extract was subjected to gel filtration, only the activity with a molecular weight of 1000 was significantly inhibited by anti-human MCF-1 mAb (7-3 mAb). Both MCA and TNF activity were detected in skin extracts from DTH inflammation sites where the specific antigen was injected. Both activities were produced at the early DTH reaction stage. In contrast, MCF-1 was produced at the late DTH reaction stage. The erythema was suppressed significantly either by anti-guinea pig TNF Ab or by 7-3 mAb, suggesting the roles of these two cytokines in a DTH reaction.

Antitumor effect of a Coliolus preparation, PSK: induction of macrophage chemotactic factor (MCF) in spleens of tumor bearing mice.

The effect of administration of PSK (Polysaccharide Kureha), a Coliolus preparation, in Meth-A solid tumors was analyzed in BALB/c mice. Spleen cells prepared from normal, non-treated Meth-A bearing, PSK-treated normal and PSK-treated tumor bearing mice were examined for induction of macrophage chemotatic factor (MCF). Only spleen cells from the latter mice produced MCF after 48 hrs of cultivation in the presence of Meth-A cells or concanavalin A (Con A). MCF-producing cells were indicated to be Lyt-1 positive, L3T4 positive and Lyt-2 negative cells in the negative elimination assay. There were no differences in the production of other cytokines including interleukin-2, interferon and tumor necrosing factor, spleen cells obtained other different groups of mice. The antitumor effect of either crude or purified MCF (molecular weight 100,000) was examined by daily consecutive intratumoral injections into Meth-A tumor tissues, and a significant inhibitory effect was detected.

A protective role of gamma/delta T cells in primary infection with Listeria monocytogenes in mice.

We have previously reported that T cells bearing T cell receptors (TCRs) of gamma/delta type appear at a relatively early stage of primary infection with Listeria monocytogenes in mice. To characterize the early-appearing gamma/delta T cells during listeriosis, we analyzed the specificity and cytokine production of the gamma/delta T cells in the peritoneal cavity in mice inoculated intraperitoneally with a sublethal dose of L. monocytogenes. The early-appearing gamma/delta T cells, most of which were of CD4-CD8- phenotype, proliferated and secreted IFN-gamma and macrophage chemotactic factor in response to purified protein derivative from Mycobacterium tuberculosis, or recombinant 65-kD heat-shock protein derived from M. bovis but not to heat-killed Listeria. To further elucidate the potential role of the gamma/delta T cells in the host-defense mechanism against primary infection with Listeria, we examined the effects of in vivo administration of monoclonal antibodies (mAbs) against TCR-gamma/delta or TCR-alpha/beta on the bacterial eradication in mice infected with Listeria. Most of alpha/beta T cells or gamma/delta T cells were depleted in the peripheral lymphoid organs at least for 12 d after an intraperitoneal injection of 200 micrograms TCR-alpha/beta mAb or 200 micrograms TCR-gamma/delta mAb, respectively. An exaggerated bacterial multiplication was evident at the early stage of listerial infection in the gamma/delta T cells-depleted mice, whereas the alpha/beta T cell-depleted mice exhibited much the same resistance level as the control mice at this stage although the resistance was severely impaired at the late stage after listerial infection.(ABSTRACT TRUNCATED AT 250 WORDS)

Antitumor Effector Mechanism of Interleukin‐lβ at a Distant Site in the Double Grafted Tumor System

Recombinant human interleukin‐lβ (IL‐lβ) Inhibited the growth of not only the right, but also the left non‐treated tumor in a double grafted tumor system. Since the antitumor activity of IL‐lβ against the right and left tumors was not seen in nude mice, lymphocytes have a key role in the antitumor effect of intratumoral administration of IL‐lβ. TIL (tumor‐infiltrating leukocytes) obtained from left and right side tumors treated with IL‐1β were examined by Winn assay for their antitumor activity against Meth‐A sarcoma in BALB/c mice. TIL from the right side clearly inhibited the growth of admixed Meth‐A cells, but control TIL did not. Spleen cells and right and left regional lymph node cells prepared from IL‐1‐treated mice were examined for Lyt‐1, Lyt‐2 and L3T4 phenotypes. The number of Lyt‐1‐positive lymphocytes increased in the spleen and in the right regional lymph nodes after intratumoral administration of IL‐1. Isolated tumor cells obtained from the right tumor treated with IL‐lβ and the left side tumor on day 6 were cultured in RPMI1640 with 10% fetal calf serum for 24 h. The culture supernatants were harvested and tested for the presence of chemotactic activity for neutrophils or macrophages. Significant neutrophil chemotactic factor and macrophage chemotactic factor activities were detected in the culture media from IL‐1‐treated tumor tissues cultured for 24 h. Neither significant neutrophil nor macrophage chemotactic activity was detected in the media from untreated tumor tissues. These results suggest that intratumoral administration of IL‐1 first induces neutrophils and macrophages in the right tumor, then Lyt‐1‐positive cells in the right regional lymph nodes and in the spleen, and subsequently induces macrophages in the left, non‐treated tumor.

Dissociated development of T cells mediating delayed-type hypersensitivity and protective T cells against Listeria monocytogenes and their functional difference in lymphokine production

CD4+ T cells mediating both delayed-type hypersensitivity (DTH) and acquired cellular resistance (ACR) were generated in mice after immunization with viable Listeria monocytogenes. In contrast, CD4+ T cells from mice immunized with killed L. monocytogenes in complete Freund's adjuvant were capable of mediating only DTH but not ACR. To determine the functional difference between T cells mediating DTH and T cells mediating ACR, we examined two different populations of T cells for profiles of lymphokine production after stimulation with a specific antigen in vitro. The production of interleukin-2 (IL-2) and IL-3 but not IL-4 was observed in both T cells mediating only DTH and those mediating DTH and ACR. In this respect, both types of T cells could be categorized into the TH1 population, and they produced macrophage chemotactic factor equally well. However, the production of gamma interferon (IFN-gamma) was observed only in T cells capable of mediating both DTH and ACR. This result was confirmed not only by an enzyme immunoassay specific for murine IFN-gamma but also by Northern (RNA) analysis for the detection of IFN-gamma mRNA. These results suggested that the TH1 population may be subdivided further into two distinct subsets and that the ineffectiveness of the killed bacterial vaccine may be partly explained by the dissociated development of T cell function.

Identification of two neutrophil chemotactic peptides produced by porcine alveolar macrophages

We have purified to homogeneity two distinct 10-kDa proteins with potent chemotactic activity for neutrophils from porcine alveolar macrophages incubated for 24 h with Escherichia coli endotoxin (lipopolysaccharide (LPS), 10 micrograms/ml). Neutrophil chemotactic activity in alveolar macrophage supernatants was concentrated by adsorption to SP-Sephadex, and purified by cation exchange and reversed phase high performance liquid chromatography. The first peptide, alveolar macrophage chemotactic factor (AMCF)-I, had chemotactic activity for both porcine and human neutrophils. The chemotactic activity for porcine neutrophils was detectable at 3 x 10(-10) M, peaked at 3 x 10(-8) M, and was comparable to that of zymosan-activated porcine serum. Segmental instillation of AMCF-I into porcine lungs caused marked neutrophil accumulation at 4 h in both bronchoalveolar lavage fluid and in lung tissue. The second peptide, AMCF-II, was active at 1.4 x 10(-9) M for porcine neutrophils, but it was less active for human polymorphonuclear neutrophils than was AMCF-I. Oligonucleotide probes to regions of the N-terminal sequences of AMCF-I and AMCF-II hybridized to mRNA recovered from LPS-stimulated alveolar macrophages. The N-terminal sequences and amino acid compositions indicate that AMCF-I and AMCF-II are distinct proteins, but that both have homologies with a family of peptide chemoattractants produced by human blood monocytes and platelets. Thus, alveolar macrophages stimulated with LPS produce two distinct 10-kDa cytokines with potent chemotactic activity for neutrophils. This indicates that there are two different peptide pathways by which alveolar macrophages can recruit neutrophils into the lung.

Macrophage chemotactic factor partially purified from granulomatous inflammation

Pathophysiologieal roles of macrophage chemotactic factor (MCF) in granulomatous inflammation were investigated. MCF was extracted in 10 mM phosphate-buffered saline, pH 7.4, from experimentally produced epithelioid cell granulomas in the liver and skin of mice. MCF activity reached a peak in the lesions prior to the time when granulomatous inflammation became maximal. MCFwas then purified from 10-week-old hepatic granulomas and 2-week-old sk in lesions by gel filtration, ion exchange column chromatography, and HPLC gel filtration. MCF from either liver or skin had a molecular weight about 650 kDa. MCF from hepatic granulomas was coupled to Affi-Gel beads and transplanted subcutaneously into naive mice. In vivo macrophage chemotaxis was observed around the beads and the cells formed a sheet, but organization of macrophages into granulomas did not occur with the MCF-active fractions. Macrophage chemotaxis alone is insufficient to elicit granulomatous inflammation.

Antitumor Effect of PSK at a Distant Site: Inductions of Interleukin‐8‐like Factor and Macrophage Chemotactic Factor in Murine Tumor

The antitumor effect of PSK, a Coriolus preparation, at a distant site was analyzed with the use of a double grafted tumor system in which male BALB/c mice received simultaneous intradermal inoculations of Meth‐A tumor in the right (106 cells) and the left (2 × 105 cells) flanks and were then injected with PSK in the right tumor on the third day thereafter. The antitumor effect of intratumoral administration of PSK in the right tumor on days 3, 4 and 5 was compared with the effect of surgical resection of the right tumor on day 5. Three out of 8 mice given PSK intratumorally became tumor‐free whereas no mouse tumor‐free in the left flank was found among the surgically resected mice. As regards sinecomitant immunity, tumor inoculation into the right flank followed by intratumoral administration of PSK on days 3 and 5 and surgical excision of the primary tumor on day 6 resulted in complete rejection of a tumor challenge in the left flank on day 21. The combination of presurgical intratumoral injections of PSK (more than 2 times) and postoperative oral administration of PSK appeared to be most effective in eradicating secondary tumors. Isolated TILs (tumor‐infiltrating lymphocytes), obtained from the right tumor (treated with PSK) and the left tumor on day 10 in the double grafted tumor system were cultured in RPMI1640 with 10% fetal calf serum for 24 h. The culture supernatants were harvested and tested for the presence of chemotactic activity for neutrophils or macrophages. Significant neutrophil chemotactic factor (NCF) and macrophage chemotactic factor (MCF) activities were detected in the culture media from PSK‐treated TILs that had been cultured for 24 h. Neither significant neutrophil nor macrophage chemotactic activity was detected in the media from untreated TILs. NCF and MCF activities were also detected in the culture supernatant from PSK‐treated tumor tissue on day 6. PSK‐induced NCF in the murine tumor was neutralized by treatment with anti‐human IL‐8 IgG, and might be murine IL‐8‐like factor. Therefore, neutrophil and macrophage infiltrations of tumors following intratumoral injections of PSK are probably mediated by inductions of IL‐8‐like factor and MCF.

Interleukin-1-induced promotion of T-cell differentiation in mice immunized with killed Listeria monocytogenes

We studied the effects of administration of recombinant interleukin-1 alpha (rIL-1 alpha) to mice after immunization with killed Listeria monocytogenes cells on the promotion of the functional differentiation of T cells in vivo. Mice immunized with killed L. monocytogenes were unable to express cell-mediated immunity to specific antigen in vivo, as determined by delayed-type hypersensitivity (DTH) and acquired cellular resistance (ACR), and splenic T cells obtained from such mice were unable to respond to rIL-2 and specific antigen and to produce IL-2 after antigenic restimulation in vitro. When rIL-1 alpha was given to mice after immunization with killed bacteria. T cells became capable of responding to rIL-2 and specific antigen in vitro. These functions of T cells were similar to those from mice immunized with viable listeriae. Moreover, using a local passive transfer system, it was found that effector T cells mediating DTH but not ACR to L. monocytogenes were generated in mice treated with rIL-1 alpha after immunization with killed bacteria. These T cells were able to produce macrophage chemotactic factor but not macrophage-activating factor or gamma interferon in vitro in response to stimulation with specific antigen. These results suggest that in vivo administration of rIL-1 alpha facilitates the maturation of antigen-specific T cells mediating DTH and that different effector T cells mediating DTH or ACR are involved in cell-mediated immunity to L. monocytogenes.

Macrophage chemotactic factor (MCF) produced by a human T cell hybridoma clone

A human T cell hybridoma clone, D6-18, producing high levels of macrophage chemotactic factor (MCF) was established by the emetine-actinomycin D selection method. MCF was found to be present not only in the culture medium but also in the cell lysate of D6-18 cells. The secretion of the MCF from D6-18 cells was effectively inhibited by disodium cromoglycate, which is an inhibitor of the degranulation of mast cells, suggesting that MCF is stored in granules. The MCF of D6-18 cells was purified from the sonicated cell lysate by ion-exchange chromatographies and high-performance liquid chromatography. The amino acid sequence of the purified MCF was revealed to be WLGREDGSE or WLGRQDGSE. The synthetic peptide WLGREDGSE showed chemotactic activity against guinea pig macrophages and human monocytes at the concentration of about 10 −8 M.