Antigen‐initiated B Lymphocyte Differentiation. Characterisation of the Primary and Secondary Immune Responses of Normal and Athymic Mice to the Hapten 4‐hydroxy‐3‐iodo‐5‐nitrophenylacetic Acid Presented on the Carrier Polymerised Bacterial Flagellin

Schlegel, RA

doi:10.1038/icb.1974.8

Cited by 17 publications

(14 citation statements)

References 9 publications

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“…CBA (H-2k) mice were immunized with 4-hydroxy-3-iodo-5-nitrophenyl acetic acid conjugated to polymerized flagellin (NIP-POL) as previously described (Schlegel, 1974). After 3 or 7 days, mice were killed, their spleens removed and assayed for haemolytic plaque forming cells.…”

Section: A Antigens On Plaque Forming Cellsmentioning

confidence: 99%

Ia ANTIGENS ON MURINE LYMPHOID CELLS: DISTRIBUTION, SURFACE MOVEMENT AND PARTIAL CHARACTERIZATION*

Goding

Nossal

Shreffler

et al. 1975

Int J Immunogenetics

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SUMMARY Antisera directed against the I (immune response gene) segment of the murine major histocompatibility complex were prepared, and the distribution of lymphoid cells binding the antisera studied by radioautography. A clear bimodal distribution of grain counts was observed in lymph node and spleen. Bone marrow lymphocytes showed heavy but heterogeneous labelling, while thymocytes were lightly but definitely labelled. Approximately 30% of lymph node cells and 60% of spleen cells were heavily labelled. The heavily labelled cells were shown to be B cells. Ia antigens are present on plaque forming cells, but probably in smaller amounts than on resting B cells. One plasma cell tumour was studied, and was found to possess little or no Ia antigen. There was no demonstrable labelling of thoracic duct lymphocytes from preparations of T cells activated to histocompatibility antigens. However, several T cell lymphomas probably possess Ia antigens, since they were rejected when transplanted into mice differing at the I region. Overall, the data indicate that Ia antigens are expressed on at least some T cells. The Ia antigens were shown to move independently of surface immunoglobulin on B cells, demonstrating the non‐identity of these molecules. Pretreatment of B cells with anti‐immunoglobulin did not inhibit binding of anti‐Ia sera. The Ia antigens could be capped by a sandwich technique in 75% of labelled cells. The Ia antigens were specifically precipitated from lysates of surface radioiodinated spleen cells, and the products analysed by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate. A single specific peak of radioactivity corresponding to a molecular weight of approximately 30,000 was found. The significance of these results in relation to immune response genes, receptors for antigen on T cells, and receptors for the Fc portion of IgG on B cells, is discussed.

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Section: A Antigens On Plaque Forming Cellsmentioning

confidence: 99%

Ia ANTIGENS ON MURINE LYMPHOID CELLS: DISTRIBUTION, SURFACE MOVEMENT AND PARTIAL CHARACTERIZATION*

Goding

Nossal

Shreffler

et al. 1975

Int J Immunogenetics

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“…Both techniques were needed to give an estimate of cell size (14). These methods were used in conjunction with an adoptive immune assay of IgM AFC progenitors reactive to the hapten 4hydroxy-3-iodo-5-nitrophenylacetic acid (NIP) conjugated to the carrier polymerized bacterial flagellin (POL) (11,(15)(16)(17). Previous studies have indicated that the CBA mouse is in a relatively unprimed state with respect to this hapten (11,15,16).…”

mentioning

confidence: 99%

“…These methods were used in conjunction with an adoptive immune assay of IgM AFC progenitors reactive to the hapten 4hydroxy-3-iodo-5-nitrophenylacetic acid (NIP) conjugated to the carrier polymerized bacterial flagellin (POL) (11,(15)(16)(17). Previous studies have indicated that the CBA mouse is in a relatively unprimed state with respect to this hapten (11,15,16). The assay allows a direct measurement of AFC progenitor function, under conditions where AFC production bears a direct, linear, arithmetic relationship to the number of progenitors transferred (17).…”

mentioning

confidence: 99%

Antigen-initiated B-lymphocyte differentiation. VIII. Sedimentation velocity and buoyant density characterization of virgin antibody-forming cell progenitors in the adoptive immune response of unprimed CBA mice to 4-hydroxy-3-iodo-5-nitrophenylacetic acid-polymerized bacterial flagellin antigen.

Fidler¹,

Howard²,

Shortman³

1976

The Journal of Experimental Medicine

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The characteristics of antibody-forming cell (AFC) progenitors lacking previous contact with specific antigen (virgin AFC progenitors) has been studied using sedimentation velocity and buoyant density separation for the investigation of physically distinct B-cell subpopulations. Functional characterization of isolated subsets was made using a quantitative adoptive immune assay for the IgM AFC progenitors responding to the antigen 4-hydroxy-3-iodo-5-nitrophenylacetic acid conjugated polymerized bacterial flagellin. Extensive heterogeneity is present among B lymphocytes, only some subpopulations of which exhibit AFC progenitor function. In the spleen of adult conventional CBA mice, atypically fast sedimenting cells of low buoyant density are active, while typical small B lymphocytes do not appear to be progenitors of IgM AFC. Spleen of adult specific pathogen-free (SPF), germfree, and athymic nude mice give similar results, although a minor population of typical slowly sedimenting dense cells are active in the latter two sources. Adult conventional bone marrow cells are as physically and functionally heterogeneous as splenic B cells, and although a significant proportion of AFC progenitor activity is found among dense, slowly sedimenting cells, most of the activity is among low density, faster sedimenting cells. In contrast to this situation in adult animals, where most of the unprimed AFC progenitors are large, atypical B cells, the spleens of neonatal mice provide a site where virgin AFC progenitors with the physical properties of typical small B lymphocytes are found. While being present in conventional and SPF neonatal spleens, these virgin cells are predominant in 7-day-old germfree mouse spleen. These findings suggest that the newborn virgin B cell is a typical small lymphocyte. However, few cells of this type are found in the adult animal. The unprimed AFC-progenitor population in the adult consists of large, fast sedimenting, low buoyant density, adherent cells, the physical properties of which are characteristic of activated B lymphocytes. It is suggested that these atypical cells are derived from the small newborn virgin B cell by the nonspecific effects of environmental antigenic stimuli.

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“…The splenic AFC response of CBA mice to an antigen such as NIP-POL (nitroiodo phenol hapten and polymerised bacterial flagellin carrier) char-13" 196 NOSSAL ET AL. , acteristicaily shows two i>eaks, around day 3-4 and day 8 (Schlegel 1974), as shown in Figure 1. This recalls the cycling AFC production kinetics reported in several systems (reviewed by Weigle 1975), but we believe in this case the two peaks derive not from oscillating feedback effects, but from two separate, sequentially related progenitor populations.…”

Section: Experimental Evidence For the Pre-progenitor -Direct Afc Promentioning

confidence: 89%

Current Problem Areas in the Study of B Lymphocyte Differentiation

Nossal¹,

Shortman²,

Howard³

et al. 1977

Immunological Reviews

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If the understandable excitement about the presence and functional significance of IgD receptors on the surface of a large percentage of B lymphocytes is to be placed in suitable historic perspective, it can only be in the context of the wider field of B lymphocyte differentiation. Knowledge about the many life stages, both pre-and post-antigenie, during which B cell development can be observed has accumulated rapidly, chiefly because of improvements in a variety of tissue culture and cloning assays and the increasing availability of cell surface markers and biophysical separation techniques to help in the identification of various B cell subsets. As is common in an emerging field, overall understanding is hampered by competing nomenclatures, and by the fact that most of the leading laboratories use differing operational criteria to describe, enumerate and assess the function of the various differentiation stages. The object of this paper is to present an overview of B cell differentiation from the viewpoint of a group of linked laboratories that has been investigating various facets of the problem for the past two decades. The analysis will reveal that the number of identifiably different cell sets which need to be understood is surprisingly large. Special emphasis will be placed on a phase of non-specific B cell activation that has not yet been the object of extensive study outside the Hall Institute., and on mechanisms of tolerance induction. We hope to show that despite the undoubted complexity of the problem, a definitive framework for classification and further research is emerging. The chief objects of the review are to suggest a provisional nomenclature and to highlight problems that are amenable to experimental approach in the near to medium term.The process of B cell differentiation, as in the case of any c^ll system which exhibits self-renewal in the mature animal, must be viewed both from the standpoint of ontogeny and from that of lymphocyte formation and proliferation in the adult. Wherever possible, the nomenclature used will 183 NOSSAL ET AL.

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Antigen‐initiated B Lymphocyte Differentiation. Characterisation of the Primary and Secondary Immune Responses of Normal and Athymic Mice to the Hapten 4‐hydroxy‐3‐iodo‐5‐nitrophenylacetic Acid Presented on the Carrier Polymerised Bacterial Flagellin

Cited by 17 publications

References 9 publications

Ia ANTIGENS ON MURINE LYMPHOID CELLS: DISTRIBUTION, SURFACE MOVEMENT AND PARTIAL CHARACTERIZATION*

Ia ANTIGENS ON MURINE LYMPHOID CELLS: DISTRIBUTION, SURFACE MOVEMENT AND PARTIAL CHARACTERIZATION*

Antigen-initiated B-lymphocyte differentiation. VIII. Sedimentation velocity and buoyant density characterization of virgin antibody-forming cell progenitors in the adoptive immune response of unprimed CBA mice to 4-hydroxy-3-iodo-5-nitrophenylacetic acid-polymerized bacterial flagellin antigen.

Current Problem Areas in the Study of B Lymphocyte Differentiation

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