<i>In vitro</i> antibody response of primed rabbit peripheral blood lymphocytes to Group A variant streptococcal polysaccharide

Read, Stanley; Braun, Dietmar

doi:10.1002/eji.1830040607

Cited by 22 publications

(17 citation statements)

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“…In a previous study it was shown that de novo synthesized anti-Av-CHO antibody can be identified in culture supernates by immunoelectrophoresis and isoelectric focusing analysis (2). In the present paper the amount of antibody produced in 1-ml cultures was measured by a modified Farr technique (10).…”

Section: Clonotype 2 Analysis Of Antibody Produced In Microculturesmentioning

confidence: 78%

“…Discussion It was the purpose of this study to investigate, in an in vitro system, the state of immunological memory of rabbits that had previously been hyperimmunized against streptococcal group polysaccharides. Previous work revealed a longlived memory persisting throughout the lifetime of the rabbit at the level of specific IgG clonotypes (1) which could also be triggered in vitro (2).…”

Section: Clonotype 2 Analysis Of Antibody Produced In Microculturesmentioning

confidence: 99%

“…For labeling with the isoptopes 125I and ~31I a prior tyramination was required to prepare the following reagents reactive with specific antibody: [~3]I]A-CHO tyr, [~25I]Av-CHO tyr (10). For coating of sheep red blood cells (SRBC) the O-stearoyl esters of the A-CHO and the Av-CHO were used (2). Both forms of SRBC were strongly agglutinated by specific antibodies and specific lysis occurred upon the addition of complement.…”

mentioning

confidence: 99%

“…Specific PFC in 1-ml cultures and microcultures were identified using the method of hemolysis in agar, as originally described by Jerne et al (11), and modified for microscope slides by Mishell and Dutton (12). The indicator systems were SRBC coated with A-CHO and Av-CHO antigens (A-CHO-O-St-and Av-CHO-O-St-SRBC) as previously described (2). Background PFC were determined with uncoated SRBC.…”

mentioning

confidence: 99%

“…Background PFC were determined with uncoated SRBC. The presence of antibody in the microcultures supernates was detected with the spot test (4) using Av-CHO-O-St-SRBC (2). In this case controls were performed also with noncoated SRBC.…”

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confidence: 99%

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Antibody response of rabbit blood lymphocytes in vitro. Kinetics, clone size, and clonotype analysis in response to streptococcal group polysaccharide antigens.

Braun¹,

Quintáns²,

Luzzati³

et al. 1976

The Journal of Experimental Medicine

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Hyperimmunization of rabbits and mice against streptococcal groups A-variant and A polysaccharides (Av-CHO and A-CHOP leads to the induction of a long-lived immunological memory. The number of clonotypes of specific IgG antibodies remains constant with time and repeated immunizations. This pattern is already stable from the moment when IgG antibodies are first detectable (1).It was previously shown that peripheral blood lymphocytes (PBL) from rabbits primed in vivo with the Av-CHO can be restimulated in vitro. It was observed that the pattern of IgG antibodies overlapped with the clonotype patterns induced in vivo (2). In the present paper the state of immunological memory was investigated by using both conventional and microculture techniques (2-4). Materials and MethodsRabbits. The rabbits used within this study were taken from a colony bred selectively for restricted high responders (5). They had been previously immunized with dead vaccines of Group A-variant (strain A486 variant, M-) streptococci and produced 10-25 mg/ml of antibody at the peak of the response. Rabbit K19-205 was additionally immunized with a Group A (strain J17A4) streptococcal vaccine 6 mo after the irdection course with the Group A-variant vaccine.Cultures. Rabbit blood was collected from the central artery of the ear into heparinized bottles. The erythrocytes were sedimented in 1% pig skin gelatin, as described by Sell and Gel] (6), and the leukocytes were washed and resuspended in RPMI 1640 medium (Microbiological Associates, Bethesda, Md.) containing 10% fetal calf serum (Rehatuin, Rebels Chemical Co., Chicago, Ill., Batch No. H73212), 5 × 10 -5 M 2-mercaptoethanol, and 50 U/ml of both penicillin and streptomycin (Microbiological Associates). Streptococcal A-variant or A vaccines were added as antigen in doses of 2.5-3 ~g rhamnose per ml (2). 1-ml cultures containing 4 × 10 ' or 2 × 10 '; cells were distributed in plastic tubes (Falcon Plastics, Los Angeles, Calif., type 2003

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Section: Clonotype 2 Analysis Of Antibody Produced In Microculturesmentioning

confidence: 78%

Section: Clonotype 2 Analysis Of Antibody Produced In Microculturesmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Antibody response of rabbit blood lymphocytes in vitro. Kinetics, clone size, and clonotype analysis in response to streptococcal group polysaccharide antigens.

Braun¹,

Quintáns²,

Luzzati³

et al. 1976

The Journal of Experimental Medicine

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Suppressor cells in rabbit peripheral blood

Luzzati¹,

Lafleur²

1976

Eur J Immunol

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Rabbit peripheral blood leukocytes (PBL) added to cultures of autologous spleen cells, primed in vivo to sheep red cells, are able to suppress with high efficacy the secondary in vitro response of the spleen cells to that antigen. Removal of nylon wool adherent cells from PBL abolishes the suppressive effect. When the PBL are fractionated by velocity sedimentation the suppressor cells can be separated from the responding cells. The circulating lymphocytes, freed from the inhibiting effect, either by nylon wool absorption or by velocity sedimentation fractionation, are able to give a strong secondary in vitro anti-SRC response, in which the long latency period, usually observed when PBL are stimulated with antigen in culture, is abolished or at least reduced. The suppressor effect present in the PBL is not due to granulocytes, platelets or erythrocytes.

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Histamine‐binding suppressor T cells in rabbit peripheral blood

1980

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Peripheral blood leukocytes (PBL) from primed rabbits were able to suppress the in vitro anti-sheep red blood cell (SRBC) plaque-forming cell (PFC) response of autologous spleen cells. A population containing the suppressor cells could be isolated from PBL by cell fractionation on columns of insolubilized histamine. In contrast to spleen cells, PBL generatd a weak secondary anti-SRBC response in vitro. A strong response was obtained with PBL freed from histamine-binding (H+) cells. The addition of these H+ cells to cultures of H-PBL caused strong suppression. The H+ suppressor cell was further characterized as a radioresistant T cell. Low-dose irradiation of H- cells resulted in a supplementary enhanced PFC response suggesting that PBL also contain a radiosensitive regulator cell which is not histamine binding.

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In vitro antibody response of primed rabbit peripheral blood lymphocytes to Group A variant streptococcal polysaccharide

Cited by 22 publications

References 17 publications

Antibody response of rabbit blood lymphocytes in vitro. Kinetics, clone size, and clonotype analysis in response to streptococcal group polysaccharide antigens.

Antibody response of rabbit blood lymphocytes in vitro. Kinetics, clone size, and clonotype analysis in response to streptococcal group polysaccharide antigens.

Suppressor cells in rabbit peripheral blood

Histamine‐binding suppressor T cells in rabbit peripheral blood

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