Limiting dilution analysis of the cells of immune system I. The clonal basis of the immune response

Lefkovits, Ivan; Waldmann, Herman

doi:10.1016/0167-5699(84)90137-3

Cited by 213 publications

(132 citation statements)

References 8 publications

Supporting

Mentioning

130

Contrasting

Order By: Relevance

“…Poisson analysis (20,21) was used to determine the frequency of immunoglobulin-secreting precursors and the maximum likelihood estimator for linear regression analysis.…”

Section: Methodsmentioning

confidence: 99%

Memory cell generation ablated by soluble protein antigen by means of effects on T- and B-lymphocyte compartments.

Karvelas

Nossal

1992

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Adult C57BL/6 mice were injected with 100 ,ug of soluble, freshly deaggregated human serum albumin (HSA) to produce partial immunologic tolerance. UnhEjected normal control (N) mice contain only 100 B cells in their spleens with the capacity to (i) be activated in vitro into clonal proliferation by Escherichia coli lipopolysaccharide plus interleukins 2, 4, and 5, (ii) form IgG1 as well as IgM antibody, and (iii) display specificity for HSA when only IgG1 is allowed to score in an enzyme-linked immunosorbent assay (ELISA). Such N mice generate ==50,000 clonable anti-HSA IgG1 antibody-forming cell precursors in their spleens after T-dependent immunization with HSA adsorbed onto alum and given with BordeteUa pertussis adjuvant. Mice prei'jected with soluble HSA (TOL)-generate far fewer anti-HSA IgG1 antibodyforming cell precursors, termed anti-HSA memory cells. Splenocytes were transferred from N or TOL mice into lethally irradiated syngeneic recipients together with syngeneic bone marrow. Whereas N splenocytes generated plentiful memory cells within 2 weeks in antigenically challenged recipients, TOL splenocytes did not. Work with Ly-5 congenic mice ruled out memory cell generation from either the host or the bone marrow inoculum within this limited time. N T cells plus TOL B cells showed consistently lowered memory cell generation. TOL T cells plus N B cells showed an even greater lowering of adoptive memory cell generation. Thus the lowered response capacity of TOL mice resided in the T-and B-cell compartments. Attempts to show a suppressor component within the T-cell population were inconclusive, but a profound defect in capacity to respond to HSA in vitro was exhibited by the CD4' T cells of TOL mice. B lymphocytes were harvested from T-dependently immunized mice 5 days after challenge, incubated with soluble HSA for 18 hr, and then adoptively transferred together with N T cells. The recently activated B cells were not rendered tolerant by this manipulation. The results argue for a major T-cell component in the process whereby soluble protein antigens ablate affinity maturation and memory cell generation.

show abstract

“…Poisson analysis (20,21) was used to determine the frequency of immunoglobulin-secreting precursors and the maximum likelihood estimator for linear regression analysis.…”

Section: Methodsmentioning

confidence: 99%

Memory cell generation ablated by soluble protein antigen by means of effects on T- and B-lymphocyte compartments.

Karvelas

Nossal

1992

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…32 The CAFC frequency was computed by means of minimization of chi by regression to the cell number at which 37% of wells were negative for CA formation, with 95% statistical precision. 33 …”

Section: Cobblestone Area-forming Cell Assaysmentioning

confidence: 99%

The relative quiescence of hematopoietic stem cells in nonhuman primates

Mahmud

Devine

Weller

et al. 2001

Blood

View full text Add to dashboard Cite

Quiescence has been thought to be required for the retention of the full biological potential of pluripotent hematopoietic stem cells (PHSCs). This hypothesis has been challenged recently by the observation that all murine PHSCs cycle continuously and constantly contribute to steadystate blood cell production. It was asked whether these observations could be extrapolated to describe hematopoiesis in higher mammals. In this series of experiments, the replicative history of PHSCs was examined in baboons by continuously administering bromodeoxyuridine (BrdU) for more than 85 weeks. The results indicate that under steady-state conditions, PHSCs remain largely quiescent but do cycle, albeit at a far lower rate than previously reported for rodent PHSCs. BrdU-labeled cycling PHSCs and progenitor cells were shown to have an extensive proliferative capacity and to contribute to blood cell production for prolonged periods of time. The proportion of PHSCs entering cell cycle could, however, be rapidly increased by the in vivo administration of granulocyte-colony stimulating factor. These data indicate that dur- IntroductionMammalian blood cell production is a tightly regulated process in which a hierarchy of cycling hematopoietic stem cell populations contributes to active hematopoiesis. [1][2][3][4][5] The formed elements of blood ultimately originate from rare pluripotent hematopoietic stem cells (PHSCs), which are capable of self-renewal and differentiation into cells belonging to each hematopoietic lineage. 6,7 PHSCs contribute to steady-state hematopoiesis and respond to a variety of stressful conditions without actually depleting their number. It is commonly believed that PHSCs remain quiescent during steady-state hematopoiesis and that stem cell quiescence plays a pivotal role in determining the self-renewal and proliferative potential of PHSCs. 8,9 In 1965, Kay 10 postulated that only a few PHSC clones actively contribute to blood cell production at any given time and that only after exhaustion of these active clones would additional PHSCs self-replicate and actively contribute to blood cell production. Experimental evidence supporting this clonal succession hypothesis was first gained by studies of the contribution of murine blood cell production by PHSCs that had been genetically modified by retroviral vectors. 11,12 The contribution of individual PHSCs to blood cell production was monitored by tracking retroviral integration sites in mature blood cells. Abkowitz and coworkers 13 also addressed this hypothesis by performing stem cell transplants in female cats who were heterozygotes for the X-linked gene glucose 6-phosphate dehydrogenase. They observed wide fluctuations in clonal contributions to hematopoiesis following initial engraftment but found that long-term reconstitution was due to a limited number of stem cell clones. 13 The validity of such reconstitution assays as an appropriate test of the clonal succession hypothesis has been questioned since the kinetics of hematopoietic reconstitution following st...

show abstract

“…No cultures contained >10 5 responder cells per well, as we found elevated levels of background proliferation (proliferation in the absence of antigen) at that cell concentration in the case of lymphocytes from rats with EAE. When the percentages of negative wells at high cell concentrations were well above the 37% cutoff level, the frequency was taken to be <1 in 150,000 cells (the limit of sensitivity of the assay) (17). Each well was supplemented with 10 U/ml IL-2 in the form of 20 µl MLA 144 supernatant 24 h after initiating the assay.…”

Section: Limiting Dilution Analysis Of Antigen-specific Cellsmentioning

confidence: 99%

Antigen-specific down-regulation of myelin basic protein-reactive T cells during spontaneous recovery from experimental autoimmune encephalomyelitis: further evidence of apoptotic deletion of autoreactive T cells in the central nervous system

1995

View full text Add to dashboard Cite

Experimental autoimmune encephalomyelitis (EAE) was induced in Lewis rats by the i.v. injection of 10 7 cloned V β 8.2 + T cells specific for the 72-89 peptide of guinea pig myelin basic protein (MBP). Some animals were injected simultaneously with 10 7 cloned T cells specific for ovalbumin (OVA). Lymphocytes were isolated from the spinal cord and from the peripheral lymphoid organs of these rats and the frequencies of MBP-peptide-specific or OVA-specific proliferating cells were estimated by limiting dilution analysis at different times after cell transfer. The frequencies of cells specific for MBP [72][73][74][75][76][77][78][79][80][81][82][83][84][85][86][87][88][89] or OVA in the spinal cord were highest 5 days after cell transfer (MBP 72-89 , 1 in 1149; OVA, 1 in 1116). On day 7, when the rats were recovering, the frequency of cells specific for MBP [72][73][74][75][76][77][78][79][80][81][82][83][84][85][86][87][88][89] in the spinal cord fell dramatically to <1 in 10 5 , while that of OVA-specific cells decreased to a much lesser extent (1 in 7001). The frequencies of MBP 72-89 -specific cells in the peripheral lymphoid organs during and after recovery were also much lower than those of OVA-specific cells. A similar pattern of downregulation of the MBP-peptide-specific, but not the OVA-specific, T cell response was observed in the spleen and mesenteric lymph nodes (MLN) of rats 38 days after the active induction of EAE by immunization with equal amounts of MBP and OVA in adjuvants. In the passively transferred model, cells isolated from the spinal cord and MLN on day 7 did not regain responsiveness to MBP 72-89 after incubation with high levels of IL-2, indicating that the unresponsiveness was not due to T cell anergy. Thus this study demonstrates that there is a specific down-regulation of the MBP 72-89 -specific T cell response during spontaneous recovery from EAE. This conclusion is consistent with our previous observation that V β 8.2 + T cells are selectively eliminated from the CNS by apoptosis during recovery from EAE induced by the passive transfer of V β 8.2 + T cells reactive to this MBP peptide. In contrast to autoreactive T cells, the non-autoreactive T cells that accumulate in the CNS during EAE appear to recirculate to the peripheral lymphoid organs.

show abstract

Limiting dilution analysis of the cells of immune system I. The clonal basis of the immune response

Cited by 213 publications

References 8 publications

Memory cell generation ablated by soluble protein antigen by means of effects on T- and B-lymphocyte compartments.

Memory cell generation ablated by soluble protein antigen by means of effects on T- and B-lymphocyte compartments.

The relative quiescence of hematopoietic stem cells in nonhuman primates

Antigen-specific down-regulation of myelin basic protein-reactive T cells during spontaneous recovery from experimental autoimmune encephalomyelitis: further evidence of apoptotic deletion of autoreactive T cells in the central nervous system

Contact Info

Product

Resources

About