摘要:

AbstractMajor basic protein and eosinophil peroxidase are predominant cationic proteins in the cytoplasmic granules of human eosinophilic leukocytes. Each of these proteins has been purified, and major basic protein and the lower molecular weight subunit of eosinophil peroxidase have been found to comigrate on polyacrylamide gel electrophoresis in sodium dodecyl sulfate with similar apparent molecular weights of about 14,700. Because previous molecular weight estimates for these proteins have not recognized the similar molecular weights of these two cationic eosinophil granule constituents, we have evaluated the possible relatedness of these proteins. Upon protein sequence analyses, it was found that the N‐terminal 20 amino acid residues of each of these two purified polypeptides differed. These findings established that major basic protein and the smaller subunit of eosinophil peroxidase, although of comparable molecular weights, are two distinct proteins within the cytoplasmic granules of human eosinophils.

ISSN:0741-5400    

DOI:10.1002/jlb.43.1.1

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

AbstractStable prostaglandin analogs are known to induce lymphopenia and neutrophilia in a dose‐dependent fashion after subcutaneous injection in rats. The purpose of the present investigation is to determine whether the prostaglandin‐induced changes in circulating leukocytes might be secondary to hypotension with the ensuing release of adrenal hormones. The adrenal medullary catecholamine epinephrine was found to induce neutrophilia in both intact and adrenalectomized rats, and the glucocorticosteroid analog dexamethasone induced a profound lymphopenia in rats as reported by previous investigators. A stable analog of PGF2α(15‐S‐15‐methyl PGF2α; M‐PGF2α) at the dose of 1 mg/kg induced marked systemic hypotension 1 h after injection, with lymphopenia and neutrophilia 6 h after injection. The non‐prostanoid hypotensive agent captopril, at a dose of 63 mg/kg, induced a hypotension of similar magnitude and kinetics to that induced by prostaglandin. Captopril also induced lymphopenia and neutrophilia at 6 h, although the neutrophilia was of lesser magnitude than that induced by prostaglandins. The prostaglandin‐induced lymphopenia was found to be mediated, at least in part, by the hypotension‐induced release of adrenal hormones, as evidenced by the abrogation of lymphopenia in prostaglandin‐treated adrenalectomized rats. Captopril‐treated adrenalectomized rats, however, did develop a significant lymphopenia, suggesting that hypotension can result in lymphopenia even in adrenalectomized rats. The M‐PGF2α‐induced neutrophilia in adrenalectomized rats, by comparison to captopril‐induced neutrophilia in adrenalectomized rats, was greater than the neutrophilia expected as the result of hypotension alone. Indeed, the M‐PGF2α‐induced neutrophilia in adrenalectomized rats was greater than the captopril‐induced neutrophilia in sham‐adrenalectomized rats. Thus, a portion of the neutrophilia induced by M‐PGF2αin intact rats may be mediated through adrenal‐independent, hemodynamic‐independent mechanisms. The possibility that M‐PGF2αmight be inducing neutrophilia via an endotoxin‐like stress reaction was investigated by examining changes in circulating white blood cells in intact and adrenalectomized C3H/HeN (endotoxin‐sensitive) and C3H/HeJ (endotoxin‐resistant) mice after prostaglandin administration. No quantitative differences in the prostaglandin‐induced neutrophilia were noted in C3H/HeJ mice as compared to the C3H/HeN mice. In conclusion, the effects of M‐PGF2αon circulating white blood cells in the intact rat are mediated in part through hemodynamic and adrenal mechanisms, and in part through adrenal‐independent, hemodynamic‐independent, and endotoxin‐like stress reaction‐independent mechanisms. Prostaglandins may contribute directly to the genesis of neutrophilia.

ISSN:0741-5400    

DOI:10.1002/jlb.43.1.5

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

AbstractAlthough the C3H/HeJ mouse is hyporesponsive to lipopolysaccharides (LPS), certain forms of the lipid A fraction have been shown to stimulate cells from this mouse strain. To determine the role of the oligosaccharide chain length on the lipid A‐induced proliferation of C3H/HeJ splenocytes, a panel of glycolipids from R‐chemotypes (Re, Rc, and Rd) and a nontoxic monophosphoryl lipid A (MPL) were tested. The MPL cells isolated from the MPL ofSalmonella minnesota, Salmonella typhlmurium, and the Reglycolipids isolated fromEscherichia coliwere found to be effective at stimulating the LPS‐hyporesponsive spleen cells. A Re‐glycolipid isolated from a different strain ofE. colicells was inactive, as were theS. minnesotaRc and Rd chemotypes. Proliferation induced by MPL and the active Re preparations was dose dependent and was inhibited by polymyxin B. Thus, if contamination of the Rc‐LPS or MPL with lipid A‐associated protein occurred, it was below functional levels. The data suggest that the C3H/HeJ spleen cells are capable of responding to certain glycolipids, but they may lack the ability to convert native LPS into a stimulatory signal. In addition, a monosaccharide precursor of lipid A (lipid X), and a monoacyl glucosamine phospholipid derivative of lipid X (MaGP), were capable of inhibiting the proliferation induced by the MPL and Re‐glycolipids. These data are compatible with the existence of a spleen cell receptor for lipid A.

ISSN:0741-5400    

DOI:10.1002/jlb.43.1.11

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

AbstractThe aim for the present studies is to examine the relationship between the phosphorylation of the 47‐kDa protein and some neutrophil responses such as degranulation, the synergistic effect of PMA on calcium ionophore‐induced degranulation, superoxide generation, and the priming of the oxidative burst produced by the chemotactic factor fMet‐Leu‐Phe and phorbol 12‐myristate 13‐acetate (PMA).Incubation of neutrophils with the protein kinase inhibitor 1‐(5‐isoquinoline‐sulfonyl)‐2‐methylplperazine (H‐7) inhibits the phosphorylation of the 47‐kDa protein produced by PMA and fMet‐Leu‐Phe but does not affect lysozyme release induced by the same stimuli or fMet‐Leu‐Phe‐induced N‐acetyl‐β‐glucosaminidase release. Furthermore, the synergistic effect of PMA on the A23187‐induced degranulation is not inhibited by H‐7. Also, pretreatment of the cells with H‐7 inhibits superoxide production produced by PMA but not by fMet‐Leu‐Phe. The inhibitory effect of H‐7 is more pronounced on the rate than on the extent of PMA‐induced superoxide release. On the other hand, N‐(2‐guanidinoethyl)‐5‐isoquinolinesulfonamide hydrochloride, HA‐1004, a less potent protein kinase inhibitor, has no Inhibitory effect on superoxide generation produced by fMet‐Leu‐Phe or PMA. In the case of superoxide production, the addition of low concentrations of PMA to rabbit neutrophils primes these cells to the subsequent stimulation by fMet‐Leu‐Phe, dramatically increasing the effect. Conversely, low concentrations of fMet‐Leu‐Phe prime the cells to PMA stimulation. The stimulation by PMA of cells primed with fMet‐Leu‐Phe is inhibited by H‐7. Moreover, the priming effect by PMA is also inhibited by H‐7. This inhibition is less pronounced at a low concentration of PMA. On the other hand, in the case of human neutrophils, the priming effect of PMA is not inhibited by H‐7. These results suggest several points. First, phosphorylation of the protein identified on two‐dimensional gel electrophoresis as having a molecular weight of 47 kDa and PI of 4.9 is not necessary for degranulation produced by either PMA or fMet‐Leu‐Phe. Second, the phosphorylation of the 47‐kDa protein is not necessary for superoxide generation, at least in the case of fMet‐Leu‐Phe and possibly for other stimuli. Third, in rabbit neutrophils, both the priming and stimulation of superoxide production with PMA are inhibited by H‐7. In human neutrophils, the priming by PMA is not affected by H‐7. In the case of human neutrophils, it is reasonable to conclude that the priming of the oxidative burst by PMA does not appear to be mediated by the phosphorylation of the 47‐kDa protein.

ISSN:0741-5400    

DOI:10.1002/jlb.43.1.18

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

AbstractIn this paper we describe the bone marrow‐derived macrophage cell line B6MP102. We describe growth characteristics, responsiveness to biological response modifiers known to activate macrophages (MPs), and the ability of B6MP102—comparable to that of peritoneal MPs—to kill and discriminate tumor cells. We demonstrate that B6MP102 is easily maintained in culture in the presence of 15% L‐M cell‐conditioned media. We have found that the B6MP102 cell line is similar to in vivo‐derived peritoneal MPs. These cells are responsive to biological response modifiers but are not spontaneously cytotoxic. They produce O−2in quantities comparable to that produced by peritoneal MPs, and they exhibit target ranges for direct killing similar to those of activated peritoneal MPs. In addition, B6MP102 is similar to MPs in mediating cytostasis. These latter two points are especially significant, suggesting that the mechanisms that MPs may use to distinguish and kill tumor cells are retained in the B6MP102 cell line. This makes the cell line a useful tool for studying MP‐tumor cell interactions.

ISSN:0741-5400    

DOI:10.1002/jlb.43.1.28

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

AbstractWe assessed the effect of protein deprivation on the ability of peritoneal macrophages from Fischer rats to produce interieukin‐1 (IL‐1) after in vitro stimulation. Pyrogenic activity of supernatants was measured by an in vivo febrile response assay. Control rats were given a 23% casein diet and protein‐malnourished rats were given an 8% casein diet for 4 weeks. IL‐1‐containing supernatants prepared from peritoneal macrophages were injected into assay rats, whose temperatures were measured for 6 hours (δT6). Rats injected with IL‐1‐containing supernatants derived from peritoneal macrophage cultures of protein‐deprived rats had significantly less fever (δT6= 0.20 ± 0.09°) than rats injected with IL‐1 containing supernatants derived from peritoneal macrophage cultures of control rats (δT6= 0.56 ± 0.09°),P<.01. Protein malnutrition leads to diminished pyrogenicity of macrophage culture supernatants and may be at least partly responsible for the decreased febrile response seen in the malnourished animals.

ISSN:0741-5400    

DOI:10.1002/jlb.43.1.36

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

AbstractFour monoclonal antibodies against the human monocyte/macrophage system, termed KI‐M1, KI‐M6, KI‐M7, and KI‐M8, are described with regard to their immunohistochemical tissue distribution pattern and their subcellular reactive sites. The differences found applying these analyses are also reflected by the various molecular weights of the recognized antigens. Based on these data it is proposed that the monocyte/macrophage system can be divided into the phagocytosing compartment on one hand and the immune accessory compartment on the other hand; the latter constitutes the interdigitating reticulum cells, the indeterminate dendritic cells, and the Langerhans cells, as well as the follicular dendritic cells (dendritic reticulum cells) as the accessory cells for T‐ and B‐cell immune response, respectively.

ISSN:0741-5400    

DOI:10.1002/jlb.43.1.41

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

AbstractWe describe a population of nonadherent cells in neonatal cord blood that, upon in vitro cultivation, develop into monocyte‐macrophages. These cells initially are negative for nonspecific esterase cytoplasmic activity, lack the monocyte marker MO.2, fall into smaller, nonmonocytic cell size areas, as determined by fluorescence‐activated cell sorter (FACS)‐assisted size analysis, and differentiate into macrophages under nonstimulatory culture conditions (in the absence of exogenous colony stimulating factors,<0.1 ng/ml endotoxin, and growth in suspension). In contrast to the adherent, committed macrophage precursors in cord blood, which differentiate into macrophages after 2‐3 days of culture, the nonadherent precursor does not acquire monocyte‐macrophage characteristics until day 14 of culture. Earlier induction is achieved by adding the monocyte‐activating agents lipopolysaccharide or 1,25 dihydroxyvitamin D3 to cultures.

ISSN:0741-5400    

DOI:10.1002/jlb.43.1.51

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

AbstractWe have previously demonstrated that granuloma macrophages from the foot pads ofMycobacterium leprae‐infected nude mice are functionally normal despite heavy intracellular burdens of bacilli. However, unlike peritoneal macrophages, these macrophages fail to restrict the intracellular growth ofToxoplasma gondiiwhen stimulated with recombinant murine gamma interferon (Mu IFN‐γ) and thus appear defective in their response to macrophage‐activating factor(s). In further characterizing this defect we have examined tumoristatic capacity, superoxide radical formation, and expression of Ia antigens on granuloma macrophages before and after treatment with Mu IFN‐γ. By all three criteria,M. leprae‐burdened granuloma macrophages failed to become activated by doses of Mu IFN‐γ that readily activate peritoneal macrophages fromM. leprae‐infected nude mice or normal Balb/c mice.M. leprae‐infected granuloma macrophages produced elevated levels of prostaglandin E2(PGE2) in vitro, which was suppressed by indomethacin. However, the inhibition of PGE2production for 48 hr in vitro did not restore normal responses to Mu IFN‐γ.

ISSN:0741-5400    

DOI:10.1002/jlb.43.1.60

出版商:Wiley    

年代:1988

数据来源: WILEY

摘要:

AbstractAdherence to the culture substratum exerted a significant influence on the growth and survival of murine bone marrow cells incubated under conditions that favored the development of mononuclear phagocytes. Bone marrow cells cultured two weeks in unprocessed polystyrene dishes, refractory to cell attachment, reached a maximum or near maximum cell density by day 11. This density was stable throughout the duration of the incubation period. Characterization of the cells on day 11 revealed an essentially pure population of mononuclear phagocytes composed of round, nonadherent cells, and slightly elongated, loosely attached cells. Bone marrow cells incubated in polystyrene tissue culture dishes processed to promote cell attachment also reached a maximum cell density by day 11. However, this maximum density was only half that attained by cells incubated in unprocessed polystyrene dishes. Furthermore, continued incubation resulted in a sharp decline in cell viability and number. The cells in tissue culture dishes on day 11 represented a pure mononuclear phagocyte population composed principally of cells adherent to the surface of the dish. The subsequent analysis of cells subcultured in processed and unprocessed polystyrene dishes indicated that adherence to the substratum modulated the growth factor requirements of bone marrow‐derived mononuclear phagocytes. Specifically, mononuclear phagocytes in tissue culture dishes expressed an elevated requirement for colony stimulating factor‐1 in order to proliferate and survive in long‐term culture.

ISSN:0741-5400    

DOI:10.1002/jlb.43.1.67

出版商:Wiley    

年代:1988

数据来源: WILEY