The chemotactic effect of mixtures of antibody and antigen on polymorphonuclear leucocytes.
01 Mar 1962-Journal of Experimental Medicine (The Rockefeller University Press)-Vol. 115, Iss: 3, pp 453-466
TL;DR: Results are described which indicate that, when antibody-antigen complexes are incubated in fresh serum, a heat-stable substance (or substances) is produced which acts directly as a chemotactic stimulus on the polymorphs.
Abstract: An in vitro technique is described for assessing the chemotactic activity of soluble substances on motile cells. Antibody-antigen mixtures when incubated (37°C) in medium containing fresh (i.e. non-inactivated) normal rabbit serum exert a strong chemotactic effect on rabbit polymorphonuclear leucocytes. Results are described which indicate that, when antibody-antigen complexes are incubated (37°C) in fresh serum, a heat-stable (56°C) substance (or substances) is produced which acts directly as a chemotactic stimulus on the polymorphs. This heat-stable chemotactic substance is not produced when antibody-antigen complexes are incubated in serum which has been heated at 56°C for 30 minutes.
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TL;DR: The progress made by lab-on-a-chip microtechnologies in recent years is analyzed, and the clinical and research areas in which they have made the greatest impact are discussed.
Abstract: Microfluidics, a technology characterized by the engineered manipulation of fluids at the submillimetre scale, has shown considerable promise for improving diagnostics and biology research. Certain properties of microfluidic technologies, such as rapid sample processing and the precise control of fluids in an assay, have made them attractive candidates to replace traditional experimental approaches. Here we analyse the progress made by lab-on-a-chip microtechnologies in recent years, and discuss the clinical and research areas in which they have made the greatest impact. We also suggest directions that biologists, engineers and clinicians can take to help this technology live up to its potential.
2,276 citations
TL;DR: In both systems the results indicate that under certain conditions leukocytes, and in particular PMNs, release into the medium a factor stimulating locomotion and exerting chemotactic action on PMNs in the vicinity.
Abstract: Polymorphonuclear leukocyte (PMN) locomotion and chemotaxis have been evaluated by direct microscopic observation of individual cells in thin slide-cover slip preparations, and also by observations on populations of cells migrating into a Millipore filter. The direct microscopic method used the polarity of the locomoting PMNs (broad, advancing lamellipodium and knoblike constriction at the rear) to record the direction of movement. The Boyden chamber Millipore assay was made more reliable by following the front of cells advancing into the filter, rather than counting the number of cells on the lower filter surface. Special modifications of the Millipore assay were necessary in order to distinguish between influences on rate of locomotion and true chemotaxis. In both systems the results indicate that under certain conditions leukocytes, and in particular PMNs, release into the medium a factor stimulating locomotion and exerting chemotactic action on PMNs in the vicinity. This cell-derived factor appears not to require serum factors for its release or action.
1,656 citations
Cites background or methods from "The chemotactic effect of mixtures ..."
...It is, of course, well known that AgGG or other factors can interact with complement to generate chemotactic factors ( 1 , 2, 4). Chemotactic factors originating from a serum source might well have masked and rendered difficult or impossible the demonstration of the cell-derived factor....
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...In recent years numerous interesting and perhaps physiologically important materials have been identified as "chemotactic" for PMNs as a result of studies made employing the Boyden Millipore system ( 1-6 )....
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...The Boyden Millipore chamber ( 1 ) offers great utility for studies on cell locomotion and chemotaxis, and most recent work has employed some modification of this system....
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TL;DR: Surprising developments suggest that in addition to leukocyte-mediated inflammation, the chemokines may also be involved in erythrocyte function and, through molecular mimicry, in microbial pathogenesis.
Abstract: Leukocytes migrate from the blood to sites of inflammation in response to locally produced chemoattractants that activate specific cell surface receptors. The primary structures of leukocyte receptors for N-formyl peptides, C5a, platelet-activating factor, and 8 of the 18 known human chemokines (interleukin-8 and related molecules) have been deduced from cloned cDNAs. All of these are seven-transmembrane-domain rhodopsin-like G protein-coupled receptors. Biochemical and molecular genetic analysis of the chemoattractant receptors indicates that the chemoattractants may have both broadly overlapping as well as specialized roles in the regulation of acute and chronic inflammation. Interestingly, the chemokine receptors have functional homologues in human cytomegalovirus and Herpesvirus saimiri. Moreover, the Duffy antigen, which mediates invasion of erythrocytes by Plasmodium vivax, a major cause of malaria, is also a chemokine binding protein. These surprising developments suggest that in addition to leukocyte-mediated inflammation, the chemokines may also be involved in erythrocyte function and, through molecular mimicry, in microbial pathogenesis.
1,296 citations
TL;DR: Macrophage migration inhibitory factor (MIF) is identified as a major secreted protein released by anterior pituitary cells in response to LPS stimulation, and it is concluded that MIF plays a central role in the toxic response to endotoxaemia and possibly septic shock.
Abstract: Cytokines are critical in the often fatal cascade of events that cause septic shock. One regulatory system that is likely to be important in controlling inflammatory responses is the neuroendocrine axis. The pituitary, for example, is ideally situated to integrate central and peripheral stimuli, and initiates the increase in systemic glucocorticoids that accompanies host stress responses. To assess further the contribution of the pituitary to systemic inflammatory processes, we examined the secretory profile of cultured pituitary cells and whole pituitaries in vivo after stimulation with bacterial lipopolysaccharide (LPS). Here we identify macrophage migration inhibitory factor (MIF) as a major secreted protein release by anterior pituitary cells in response to LPS stimulation. Serum analysis of control, hypophysectomized and T-cell-deficient (nude) mice suggests that pituitary-derived MIF contributes to circulating MIF present in the post-acute phase of endotoxaemia. Recombinant murine MIF greatly enhances lethality when co-injected with LPS and anti-MIF antibody confers full protection against lethal endotoxaemia. We conclude that MIF plays a central role in the toxic response to endotoxaemia and possibly septic shock.
990 citations
TL;DR: A 48-well chemotaxis chamber to minimize manipulation time and amount of material required by the larger blindwell or Boyden chemot axis chamber is designed, suitable for clinical research on the functional state of monocytes in large groups of patients.
Abstract: We designed a 48-well chemotaxis chamber to minimize manipulation time and amount of material required by the larger blindwell or Boyden chemotaxis chamber. Cell and chemoattractant dose-response curves showed that results were comparable to our better than those obtained with blindwell chambers. The volume of chemoattractant per well is 25 microliter; the number of cells can be as low as 10,000. The time needed for setting up this multiwell unit and for staining the membrane filter sheet is negligible. Combined with the use of an image analyzer to count the number of migrated cells, the method is suitable for clinical research on the functional state of monocytes in large groups of patients.
979 citations
References
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182 citations
"The chemotactic effect of mixtures ..." refers background in this paper
...The supernatant (1) obtained by centrifugation of the antigen-antibody precipitate after incubation for 1 hour possesses considerable chemotactic activity....
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...(1)~ 25 (2) 230 (3) 450 (4) 355 (5) 296 (6) 45 Migration...
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...The mixtures in the test compartments were as follows:-(1) 0....
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...;~ The figures in parentheses refer to the numbers of the chambers with mixtures as follows : ~ (1) 0....
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...The figures in parentheses indicate the number of the chamber as follows:-(1) 0....
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165 citations
"The chemotactic effect of mixtures ..." refers background in this paper
...(1)~ 25 (2) 230 (3) 450 (4) 355 (5) 296 (6) 45 Migration...
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...However, the supernatant (3) which followed reincubation at 37°C of the precipitate in fresh medium on the 2nd day was again active....
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...(3) Mixture C (antibody and antigen inactivated before incubation at...
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TL;DR: The results obtained suggest an explanation for the suitability of human, but not rabbit, blood for opsonophagocytic tests designed to measure type-specific streptococcal antibodies.
Abstract: Studies have been made on phagocytosis and killing of Group A streptococci during mixing with suspensions of leucocytes in vitro . Under appropriate test conditions an anti-phagocytic effect can be demonstrated for the streptococcal hyaluronic acid capsule as well as for its M protein.
The results obtained suggest an explanation for the suitability of human, but not rabbit, blood for opsonophagocytic tests designed to measure type-specific streptococcal antibodies. Human sera contain a factor which counteracts the anti-phagocytic effects of streptococcal hyaluronic acid capsules, and hence human blood serves well for detection of antibodies which combine with the only other phagocytosis-resisting component of this microorganism, namely M protein. In contrast, rabbit sera contain none of this factor, and addition of antibody to M protein to phagocytic test systems employing rabbit serum does not necessarily render the streptococci susceptible to engulfment by white cells, since the hyaluronic acid capsule may continue to interfere with phagocytosis.
The nature of the human serum factor which opsonizes encapsulated streptococci is unknown. It does not appear to be an antibody or an enzyme capable of depolymerizing hyaluronic acid.
75 citations
"The chemotactic effect of mixtures ..." refers background or methods in this paper
...(1)~ 25 (2) 230 (3) 450 (4) 355 (5) 296 (6) 45 Migration...
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...Polymorptwnuclear leucocytes were obtained from the peritoneal cavity of rabbits by the technique described by Ilirseh and Church (4)....
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...Set b (4) Normal medium (5) " " (6) " " Set c (7) Tuberculoprotein 0....
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TL;DR: It is posed to discuss whether any suggestions seems that at least in some cases normality can rom the study of the living lymphocyte only be maintained when specialized cells are in.
Abstract: ;CTORS of Churchilliana may like to be influencing endocrine organs; bu t d of the story of the pathologist who of the body and all their cells areneted. ed a blood count on him. On reviewing Perhaps indeed there is a simple mechanism ps, as the Prime Minister described the whereby the chemical composition of all cells is of the differential count, he commended kept basically uniform. In tissue culture cells rmorphs and monocytes for their merimay often be observed to sequestrate portions ervice. \"But what are these lymphocytes of their cytoplasm, and many, perhaps most cells, nophils doing?\" The pathologist admitted constantly take up from their environmental did not know-\"but we find them very fluid small droplets by the mechanism known as i the laboratory.\" \"Sir\", was the stern \"pinocytosis\". In this way the special comThe human frame was built before the ponents of one cell may be, in however small a )ry. Run away and find out what they concentration, incorporated into the substance \" So far as I know, however, these of others. In the case of many tissues \"celliave not yet been fulfilled, so difficult is it lines\" may be established and readily subLd a biological horizon. cultured in vitro. We have isolated such strains ndeed astonishing that no essential funcfrom many types of pathological thyroids includLs been ascribed to the lymphocyte. ing carcinoma, from malignant melanoma, ng to Osgood (1954) the human body Hodgkin's disease, seminoma of testis, and 1,300 grams of lymphocytes outside the foetal human liver. We were late-comers in this lymph nodes, and bone-marrow; and field; but in common with all others we -find '1956) estimated that their weight in the that such easily cultivated cells quickly deviate arrow was 70 grams. Osgood again from their early form to a primitive cell stimated that there were 3 grams in the indistinguishable from the HeLa cell of Gey. ng blood, and 100 grams in lymphatic Moreover in the case of the thyroid these cell The weight of the human liver is 1,600 lines cease to be able to synthesize hormones not very much more than the total in vitro. They differ from the freshly isolated )f the lymphocytes. Yet Trowell (1958) thyroid in many other ways; for example, in their y at the end of his survey of the literature reaction to certain viruses, in the rotation of the be regretfully concluded that the office nucleus, and in the manner in which one cell is 'inderella cell is obscure\". related to another (Pulvertaft et al., 1959). It pose to discuss whether any suggestions seems that at least in some cases normality can rom the study of the living lymphocyte only be maintained when specialized cells are in
41 citations
TL;DR: An egg-adapted strain of mumps virus has been cultivated in suspended cell tissue cultures and produces measurable quantities of hemagglutinin in both Influenza A and PR8 strains.
Abstract: Conclusions1. An egg-adapted strain of mumps virus has been cultivated in suspended cell tissue cultures. 2. In such media the mumps virus as well as Influenza A virus (PR8 strain) produce measurable quantities of hemagglutinin.
38 citations