scholarly journals STUDIES ON ANTIBODY FORMATION BY PERITONEAL EXUDATE CELLS IN VITRO

1960 ◽  
Vol 111 (4) ◽  
pp. 573-600 ◽  
Author(s):  
John M. McKenna ◽  
Kingsley M. Stevens
Keyword(s):  
Tissue Culture ◽  
Cell Types ◽  
Peritoneal Exudate ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Peritoneal Exudate Cells ◽  
Carbon Particles ◽  
Secondary Systems

Cells from peritoneal exudates of rabbits sacrificed 3 days after an intraperitoneal injection of sterile mineral oil were grown in tissue cultures in medium 199 (75 per cent); normal rabbit serum (25 per cent). Antibody produced by the cells was assayed by an hemagglutination technique in which the antigens used were adsorbed to formalinized tanned sheep erythrocytes. These sensitized cells agglutinate in the presence of antibody specific to the adsorbed antigen. It has been demonstrated that: Peritoneal exudate cells produced hemagglutinating antibody to bovine gamma globulin (BGG) in a replicating tissue culture system for approximately 3 weeks when taken from animals given either primary or secondary injections of BGG. The mean hemagglutinating titer was 30 for the primary and 32 for the secondary systems. Since the other cell types did not persist, it is felt that monocytes were responsible for these results. Monocytes taken from normal rabbits and exposed to either BGG or egg albumen (EA) in vitro produced titers of 28 for about 2 weeks. Monocytes taken from rabbits given hyperimmunizing injections of BGG produced titers of 147 for about 1 week. Endotoxin from Salmonella typhosa caused the monocytes to form antibody as if they had been taken from hyperimmunized rabbits. This was true both when the antigen was given in vivo together with the endotoxin as well as when the cells were exposed to antigen in vitro. The titers were 223 and 97, respectively. Neither freshly harvested nor cultured monocytes were phagocytic for carbon particles or bacteria in vitro. Monocytes in tissue culture appeared to assume the morphology of fibroblasts, but did not stain with the characteristics of fibroblasts. The morphologic changes and staining characteristics of monocytes in tissue culture have been described. The implications of these findings have been discussed and an attempt made to integrate them into general biological theory.

scholarly journals Immunogenic variants obtained by mutagenesis of mouse mastocytoma P815. II. T lymphocyte-mediated cytolysis.

1980 ◽  
Vol 152 (5) ◽  
pp. 1184-1193 ◽  
Author(s):  
T Boon ◽  
J Van Snick ◽  
A Van Pel ◽  
C Uyttenhove ◽  
M Marchand
Keyword(s):  
Tumor Cell ◽  
T Lymphocyte ◽  
Peritoneal Exudate ◽  
Antigenic Specificity ◽  
Spleen Cells ◽  
Peritoneal Exudate Cells ◽  
Specific Antigens

Tumor cell variants that were rejected by syngeneic mice (tum-) were obtained from mastocytoma P815 by mutagenesis (as described in the accompanying report (13). A considerable T lymphocyte-mediated lysis was observed upon incubation of these tum- variants with peritoneal exudate cells collected a few days after an intraperitoneal challenge of immune animals. Spleen cells from these animals were cytolytic after stimulation in vitro with the immunizing variant. New antigens, absent from the original P815 tum+ cells, were detected on 15 of the 21 tum- variants that were tested. All these antigens appeared to be different. No new antigen was detected on any of 10 mutagenized P815 clones that had retained their ability to form tumors. We compared the evidence obtained in vivo and in vitro for the presence of specific antigens on five tum- variants. Three variants were shown both in vivo and in vitro to carry an individual antigen. One showed no specificity either in vivo or in vitro. However, for one variant, no specificity was observed in vivo, although cytolysis tests demonstrated the existence of a singular antigenic specificity.

Immune responses to hapten conjugates in vitro

1975 ◽  
Vol 8 (4) ◽  
pp. 507-522
Author(s):  
Sirkka Kontiainen ◽  
O. Mäkelä ◽  
M. Hurme
Keyword(s):  
Tissue Culture ◽  
Immune Responses ◽  
Cell Types ◽  
Antibody Responses ◽  
Tissue Cultures ◽  
Cell Type ◽  
Animal Body ◽  
Do So

Several functions of the animal body can take place in cell or tissue cultures with almost unreduced efficiency and precision. Functions, where only one cell type is involved, often do so, but also some differentiation steps where interactions between two or more cell types are clearly needed can take place in tissue culture (Saxén et al. 1968).Most immune responses require collaboration between two or more cell types (Claman, Chaperon & Triplett, 1966; Miller & Mitchell, 1968; Feldmann & Nossal 1972c). Some of them can be easily induced in vitro but others cannot. Even when antibody responses can be induced in vitro their intensity varies a great deal. With some antigens and under some circumstances a response in vitro can be nearly as strong as one in vivo. A crude comparison can be derived from responses in vitro and in vivo to the same antigen, conjugate of hapten NIP and pneumococcal polysaccharide type III (NIP-SIll, Nakamura, Ray & Mäkelä, 1973).

scholarly journals Macrophage-Eosinophil Interactions in the Inflammatory Response to Trichinella spiralis

Blood ◽  
1974 ◽  
Vol 44 (1) ◽  
pp. 131-136 ◽  
Author(s):  
Ronald S. Walls ◽  
Peter Hersey ◽  
Paul G. Quie
Keyword(s):  
Inflammatory Response ◽  
Indirect Immunofluorescence ◽  
Trichinella Spiralis ◽  
Close Association ◽  
Cell Types ◽  
Tissue Reaction ◽  
Peritoneal Exudate ◽  
Peritoneal Cells

Abstract Little is understood of the reasons why eosinophils accumulate in vivo, although a tissue reaction to administered antigen has been shown to be a prerequisite for the response. The present experiments aim to understand the significance of the tissue reaction to parasitic larvae in the chain of events leading to eosinophilia. Trichinella spiralis larvae were injected intraperitoneally in rats, and the peritoneal exudate was examined at intervals. Eosinophils were closely associated with macrophages rather than with other cell types or with parasites. A striking collection of eosinophils around individual macrophages was noted 48 hr after injection, and in particular following a second challenge. Use of indirect immunofluorescence techniques showed that these coincided with presence of immunoglobulin on macrophage surfaces. Incubation of peritoneal cells in medium containing 0.25% trypsin reduced macrophage surface immunofluorescence and numbers of rosettes. Rosettes could be constituted in vitro by incubation of normal peritoneal cells in medium containing antigen and antibody. These findings suggest that in the tissue reaction to parasitic larvae a close association exists between eosinophils and macrophages.

scholarly journals Interleukin-6 mRNA and protein increase in vivo following induction of acute thrombocytopenia in mice

Blood ◽  
1991 ◽  
Vol 77 (2) ◽  
pp. 286-293 ◽  
Author(s):  
LH Cox ◽  
T Downs ◽  
K Dagg ◽  
J Henthorn ◽  
SA Burstein
Keyword(s):  
Specific Activity ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Experimental Animals ◽  
Steady State Levels ◽  
Phosphate Buffered Saline ◽  
Protein Serum ◽  
Significant Increment

Abstract Induction of experimental thrombocytopenia in rodents results in the enhancement of megakaryocytic growth and differentiation. Interleukin-3 (IL-3) and IL-6, cytokines with a broad spectrum of biologic activities, stimulate megakaryocytopoiesis in vitro. To determine if expression of these factors might increase in response to experimental thrombocytopenia, we measured steady-state levels of IL-3 and IL-6 mRNA following rabbit antiplatelet serum (APS) injection. Groups of mice were injected intravenously with 0.2 mL APS while control animals received rabbit antilymphocyte serum (ALS), normal rabbit serum (NRS), or phosphate-buffered saline (PBS). At various times up to 72 hours after injection mice were exsanguinated and splenectomized. Platelet counts in the experimental animals were less than 12% of controls. Splenic RNA was hybridized in solution to 32P-UTP-labeled cRNA probes for IL-3 and IL-6. RNase-resistant hybrids were resolved on denaturing gels and visualized autoradiographically. IL-3 hybrids were undetectable at all time points tested, irrespective of the film exposure time or specific activity of the probe. Conversely, IL-6 hybrids were easily visualized and showed peak expression at 1.5 to 2.0 hours. By 3 hours, IL-6 mRNA had returned almost to the level of the controls. Similar results were observed in the bone marrow, although maximal IL-6 mRNA in that tissue was observed 4 hours following APS administration. To determine if this mRNA increment was associated with a concomitant increase in bioactive protein, serum was tested for its ability to stimulate IL-6-dependent B9 cells. At 1.75 hours following injection, experimental animals showed a small but significant increment in IL-6 activity compared with controls (200 +/- 30 U/mL IL-6 compared with 129 +/- 17 U/mL in ALS-injected controls, 106 +/- 17 U/mL in NRS-injected controls and 84 +/- 17 U/mL in PBS-injected controls). The data show that IL-6 mRNA and bioactive protein increase in response to acute immunothrombocytopenia, while no increment in IL-3 is detectable. These results suggest that IL-6 may play a role in the physiologic response to acute immunothrombocytopenia.

STUDIES ON TISSUE CULTURE OF EQUINE OVARIAN CELL TYPES: EFFECT OF GONADOTROPHINS AND STAGE OF CYCLE ON STEROIDOGENESIS

1969 ◽  
Vol 43 (3) ◽  
pp. 415-425 ◽  
Author(s):  
CORNELIA P. CHANNING
Keyword(s):  
Tissue Culture ◽  
Cell Cultures ◽  
Granulosa Cells ◽  
Horse Serum ◽  
Cell Types ◽  
Phase Cell ◽  
Medium Change ◽  
Midluteal Phase

SUMMARY Granulosa cells were harvested from follicles of mares at various stages of the oestrous cycle and maintained in a tissue culture medium containing 15% horse serum, 30% medium '199' and 55% Hanks's solution. Between days 4 and 10 of culture the granulosa cells harvested from small follicles (1–2 cm. diam.) of mares in the midluteal phase of the cycle secreted an average of 0·36 pg. progesterone/cell/day. Cells harvested from large follicles of mares in the late and/or early oestrous stage of the cycle secreted an average of 29·5 pg. progesterone cell/day; the cells harvested from the large vascular follicles found at oestrus secreted an average of 173 pg./cell/day. The small, poorly vascularized follicles found adjacent to the large vascular follicles of mares in oestrus yielded cells which secreted less progesterone than those from the larger follicles. Addition of 5 to 10 i.u. human chorionic gonadotrophin (HCG)/ml. at each medium change (every 2–3 days) or for the first 4 days of culture brought about a marked stimulation of progesterone secretion in cultures of ' mid-luteal phase' cells which was maximal after 4 to 7 days. Pregnenolone was converted primarily to progesterone, 20α-hydroxypregn-4-en-3-one and 17-hydroxyprogesterone; the metabolism was not significantly altered by the addition of a mixture of 10 i.u. HCG plus 10 i.u. pregnant mare serum gonadotrophin (PMSG). Cells harvested from mares in oestrus converted pregnenolone to progesterone in a higher yield compared with cells harvested from mares in the midluteal phase of the cycle. Addition of 10 i.u. HCG/ml. or PMSG plus HCG (10 i.u. each/ml.) stimulated aromatization of testosterone by 'midluteal phase' cultures but not by 'oestrous phase' cell cultures. These results demonstrate that the in vivo environment as well as the in vitro conditions influence the steroidogenic activity of equine granulosa cell cultures.

scholarly journals Interleukin-6 mRNA and protein increase in vivo following induction of acute thrombocytopenia in mice

Blood ◽  
1991 ◽  
Vol 77 (2) ◽  
pp. 286-293
Author(s):  
LH Cox ◽  
T Downs ◽  
K Dagg ◽  
J Henthorn ◽  
SA Burstein
Keyword(s):  
Specific Activity ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Experimental Animals ◽  
Steady State Levels ◽  
Phosphate Buffered Saline ◽  
Protein Serum ◽  
Significant Increment

Induction of experimental thrombocytopenia in rodents results in the enhancement of megakaryocytic growth and differentiation. Interleukin-3 (IL-3) and IL-6, cytokines with a broad spectrum of biologic activities, stimulate megakaryocytopoiesis in vitro. To determine if expression of these factors might increase in response to experimental thrombocytopenia, we measured steady-state levels of IL-3 and IL-6 mRNA following rabbit antiplatelet serum (APS) injection. Groups of mice were injected intravenously with 0.2 mL APS while control animals received rabbit antilymphocyte serum (ALS), normal rabbit serum (NRS), or phosphate-buffered saline (PBS). At various times up to 72 hours after injection mice were exsanguinated and splenectomized. Platelet counts in the experimental animals were less than 12% of controls. Splenic RNA was hybridized in solution to 32P-UTP-labeled cRNA probes for IL-3 and IL-6. RNase-resistant hybrids were resolved on denaturing gels and visualized autoradiographically. IL-3 hybrids were undetectable at all time points tested, irrespective of the film exposure time or specific activity of the probe. Conversely, IL-6 hybrids were easily visualized and showed peak expression at 1.5 to 2.0 hours. By 3 hours, IL-6 mRNA had returned almost to the level of the controls. Similar results were observed in the bone marrow, although maximal IL-6 mRNA in that tissue was observed 4 hours following APS administration. To determine if this mRNA increment was associated with a concomitant increase in bioactive protein, serum was tested for its ability to stimulate IL-6-dependent B9 cells. At 1.75 hours following injection, experimental animals showed a small but significant increment in IL-6 activity compared with controls (200 +/- 30 U/mL IL-6 compared with 129 +/- 17 U/mL in ALS-injected controls, 106 +/- 17 U/mL in NRS-injected controls and 84 +/- 17 U/mL in PBS-injected controls). The data show that IL-6 mRNA and bioactive protein increase in response to acute immunothrombocytopenia, while no increment in IL-3 is detectable. These results suggest that IL-6 may play a role in the physiologic response to acute immunothrombocytopenia.

scholarly journals ANTIBODY FORMATION AGAINST TREPONEMA PALLIDUM—AGGLUTINATION

1915 ◽  
Vol 21 (6) ◽  
pp. 576-582 ◽  
Author(s):  
Hans Zinsser ◽  
Joseph Gardner Hopkins
Keyword(s):  
Quantitative Difference ◽  
Treponema Pallidum ◽  
Diagnostic Value ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Human Beings ◽  
Human Sera ◽  
Normal Human

It has been shown by our experiments that the serum of rabbits treated with emulsions of Treponema pallidum contains agglutinating substances. Normal rabbit serum also possesses agglutinating power for this organism, but, as in the case of normal bacterial agglutinins, to an extent very much inferior to that possessed by the sera of immunized animals. Normal human sera will agglutinate similar pallidum emulsions, as will the sera of certain syphilitic patients with positive Wassermann reactions. Whether or not there is a quantitative difference of diagnostic value between the sera of normal human beings and those of syphilitics remains to be seen. The sera of rabbits immunized with strain A agglutinate Noguchi's strain 9 in dilutions as high as 1 to 500. We regard as the most important result of these experiments the demonstration of definite antibodies in the circulation of animals treated with dead emulsions of Treponema pallidum. Since it is our belief that the agglutinating effect is due to an antibody essentially the same as that which produces bactericidal, precipitating, and opsonic effects, i. e., that there is probably one type of antibody only, we believe that the demonstration of agglutinins establishes the fact that in syphilis as in bacterial diseases the host responds by the formation of antibodies or sensitizers specific for the treponema. Spirocheticidal experiments with these sera, both in vitro and in vivo, are in progress.

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