A Modified Antibody Forming Cell Assay for Detecting Cells Producing a Particular Class of Antibody

Kraft, N.; Wistar, Richard

doi:10.1038/icb.1971.2

Cited by 1 publication

(5 citation statements)

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“…1 ; both the levels of AFC in blood and the serum show suggestions of similar peaks . Similar fluctuations of the immune response to this and other antigens have been observed previously in other species (3,4,26) . After day 7, there was a gradual decline in AFC numbers.…”

Section: General Kinetics Of the Responsesupporting

confidence: 89%

“…Forced feeding was employed to maintain the animals in a good nutritional state, a factor which has been found essential to obtain a normal response (19) . Under these conditions, the over-all kinetics of the toad response was similar to that seen in mice and rats (3,4), as shown in Fig . 1 .…”

Section: General Kinetics Of the Responsesupporting

confidence: 70%

“…There was a clearcut progression of both density changes and size changes in the AFC population as a function of time after antigenic stimulation . A much more complex situation, lacking this striking progression, was noted when AFC development was followed by density in the rat (17,4) or by size in the mouse (30) . In part, this has been ascribed to the restriction of the response to a single antibody class in the toad, and to a generally simpler immune apparatus in the phylogenetically more primitive animal (4) .…”

Section: Afc Development In the Toad As A Model Of Cell Dif Ferentiationmentioning

confidence: 99%

“…A much more complex situation, lacking this striking progression, was noted when AFC development was followed by density in the rat (17,4) or by size in the mouse (30) . In part, this has been ascribed to the restriction of the response to a single antibody class in the toad, and to a generally simpler immune apparatus in the phylogenetically more primitive animal (4) . The question of the relative degree of synchrony of the response in the different species may also be important .…”

Section: Afc Development In the Toad As A Model Of Cell Dif Ferentiationmentioning

confidence: 99%

“…The development of antibody-forming cells (AFC) tiated by a defined antigenic stimulus, and leading during an immune response is a particularly attrac-to the formation of a specific antibody as a defined tive model of a differentiation process, being ini-end product . The advent of assays for AFC (1)(2)(3)(4) and the development of culture systems capable of sustaining an immune response in vitro (5)(6)(7) has made possible the detailed study of AFC maturation . Such studies would be greatly facilitated if cells at various, defined stages of the response could be separated from each other.…”

Section: Introductionmentioning

confidence: 99%

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Differentiation of Antibody-Forming Cells in Toad Spleen

Kraft

Shortman

1972

The Journal of Cell Biology

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Antibody-forming cells (AFC), developing in toad spleen after stimulation with polylnerized flagellin, were studied with an immune adherence assay. Differentiation was followed by several parameters : thymidine uptake to monitor dividing cells ; equilibrium density centrifugation in albumin gradients to monitor cell density ; microscopic measurements and sedimentation velocity separation to monitor cell size ; stained preparations to follow cell morphology. Almost all AFC observed early in the response were dividing cells ; the proportion of dividing AFC dropped to 4% 2 wk after stimulation . The earliest AFC detected (3 days) formed a relatively homogeneous light density population, and were purified 17-fold by equilibrium density centrifugation . As the response developed, additional denser peaks were found, so that late in the response dense AFC predominated . Dividing AFC were confined to the light density region throughout the response . Cell diameter measurements revealed that the earliest AFC were all very large cells . In a manner analogous to the density changes, smaller AFC appeared as the response developed until they finally comprised the majority of the AFC population . Dividing AFC were always relatively large, but encompassed a wide range of sizes . Sedimentation velocity separation was employed in a closer study of the immature AFC ; they were purified 140-fold by this procedure . The earliest AFC consisted of several readily separable size populations in the range 9-18 u diameter . The presence of separate peaks related by factors of two in volume suggested that the largest cells undergo a series of halving divisions before entering a division growth cycle . The results suggest an AFC differentiation sequence from a very large, light density, dividing "blast" cell to a nondividing cell with the size, density, and morphological appearance of a small lymphocyte . Stages of this sequence can be defined and selected out for investigation, using sedimentation velocity and equilibrium density centrifugation as complementary techniques.

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Section: General Kinetics Of the Responsesupporting

confidence: 89%

Section: General Kinetics Of the Responsesupporting

confidence: 70%