Synthesis of antibody by spleen cells after exposure to kiloroentgen doses of ionizing radiation

Makinodan, Takashi; Nettesheim, Paul; Morita, Toshiteru; Chadwick, Carol J.

doi:10.1002/jcp.1040690312

Cited by 34 publications

(7 citation statements)

References 18 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…47,48 Recently, studies have shown that plasma cells can survive for more than 90 days in bone marrow and spleen in lymphocytic choriomeningitis virus-immune mice that were given 600 cGy of TBI. 49 The immune response in platelet-derived FVIII gene therapy is not our primary focus in the current study, but some experiments have been necessary to understand our results.…”

Section: Discussionmentioning

confidence: 99%

Syngeneic transplantation of hematopoietic stem cells that are genetically modified to express factor VIII in platelets restores hemostasis to hemophilia A mice with preexisting FVIII immunity

Shi

Fahs

Wilcox

et al. 2008

Blood

123

View full text Add to dashboard Cite

Although genetic induction of factor VIII (FVIII) expression in platelets can restore hemostasis in hemophilia A mice, this approach has not been studied in the clinical setting of preexisting FVIII inhibitory antibodies to determine whether such antibodies would affect therapeutic engraftment. We generated a line of transgenic mice (2bF8) that express FVIII only in platelets using the platelet-specific ␣IIb promoter and bred this 2bF8 transgene into a FVIII null background. Bone marrow (BM) from heterozygous 2bF8 transgenic (2bF8 tg؉/؊ ) mice was transplanted into immunized FVIII null mice after lethal or sublethal irradiation. After BM reconstitution, 85% of recipients survived tail clipping when the 1100-cGy (myeloablative) regimen was used, 85.7% of recipients survived when 660-cGy (nonmyeloablative) regimens were used, and 60% of recipients survived when the recipients were conditioned with 440 cGy. Our further studies showed that transplantation with 1% to 5% 2bF8 tg؉/؊ BM cells still improved hemostasis in hemophilia A mice with inhibitors. These results demonstrate that the presence of FVIII-specific immunity in recipients does not negate engraftment of 2bF8 genetically modified hematopoietic stem cells, and transplantation of these hematopoietic stem cells can efficiently restore hemostasis to hemophilic mice with preexisting inhibitory antibodies under either myeloablative or nonmyeloablative regimens. IntroductionHemophilia A is a severe congenital bleeding disorder caused by a deficiency of clotting factor VIII (FVIII). 1 Currently, hemophilia is treated with protein replacement therapy using either plasmaderived or recombinant FVIII. 2 Although FVIII replacement markedly improves the life expectancy of patients with hemophilia, up to 30% of patients with severe hemophilia A develop antibodies after FVIII replacement therapy. [3][4][5][6][7] These antibodies cause the clinical failure of treatment in response to routine replacement therapy for bleeding episodes and therefore are referred to as FVIII inhibitors. [8][9][10] Clinically, inhibitor titers greater than 5 Bethesda units (BU)/mL are considered untreatable using routine FVIII replacement. 11 Induction of immune tolerance to the FVIII protein is a treatment option for the eradication of anti-FVIII inhibitors, but it is very expensive and may not always be effective. [12][13][14] Infusion of NovoSeven (FVIIa) may also restore hemostasis in patients with inhibitors, but it is more costly because of its short half life and may result in thrombotic complications. [15][16][17] Gene therapy could be an alternative treatment for hemophilia A. There has been substantial progress in gene therapy of hemophilia A in preclinical trials. [18][19][20][21][22][23][24][25][26][27][28][29] Gene therapy of hemophilia A with preexisting FVIII immunity is especially challenging because circulating inhibitory antibodies in plasma may inactivate functional FVIII if it is constitutively secreted into plasma. Therefore, developing a mechanism for secure cellular del...

show abstract

Section: Discussionmentioning

confidence: 99%

Syngeneic transplantation of hematopoietic stem cells that are genetically modified to express factor VIII in platelets restores hemostasis to hemophilia A mice with preexisting FVIII immunity

Shi

Fahs

Wilcox

et al. 2008

Blood

123

View full text Add to dashboard Cite

show abstract

“…Even when thymidine-3H was not removed from the cultures, the dose of radiation delivered to the cells by thymidine-SH in solution would not affect antibody synthesis, as was previously reported (10). A recent report (11) has shown that very high doses (10,000 rads) of ionizing irradiation delivered to cells had no observable effect on their ability to synthesize antibody.…”

Section: Discussionmentioning

confidence: 59%

Antibody Production in Vitro

Harris

1968

The Journal of Experimental Medicine

View full text Add to dashboard Cite

The in vivo injection of antigen stimulates the proliferation of lymphoid cells and the appearance of specific antibody-producing cells (1-3), this cellular reaction being more marked during secondary responses than after primary immunization.Antibody production in vitro has been described by many investigators and has been well reviewed (4, 5), but direct correlation between cellular proliferation and the appearance of cells producing specific antibody is lacking, although studies of in vivo situations have been reported 1 (6, 7) indicating that antibodyproducing cells arose from DNA-synthesizing cells in response to antigen.The present experiments investigated the relationship between DNA synthesis and the production of antibodies to sheep erythrocytes (SRC's) in vitro by single cells from the spleens of rabbits killed after a boost injection of antigen 2 days previously. It was shown that the rate of DNA synthesis, as measured by the incorporation of radioactive thymidine, was stimulated in spleen cell suspensions from such rabbits by the presence of SRC's (8).The results of the studies reported here indicate that the major proportion of antibody-producing cells in these cell suspensions arose from the DNA-synthesizing population which was stimulated to divide by the presence of antigen, but that only a very small proportion of these proliferating cells was occupied in the production of specific hemolytic antibodies. Materials and MethodsA,imals.--New Zealand white rabbits weighing at least 2 kg were obtained from Purdey's Farm, Rochford, Essex, England.Antigen.--Sheep erythrocytes (SRC's) were supplied in Alsever solution by Burroughs Wellcome and Co., London, England.Radioactive-Labeled Materials.--Thynfxdine-~C at 30.4 mc/m~ and thymidine-SH at 2-16 c/n~ were obtained from the Radiochemical Centre, Amersham, England.

show abstract

“…This could be explained by the known radioresistance of plasma cells in comparison to other cells present in the lymphatic tissue (19,20). It is noteworthy that this background of Ig activity shows a pattern of bands, which suggests that the focal constitution of the normal lymphoid organ (1) is not completely disrupted by the radiation damage.…”

Section: Discussionmentioning

confidence: 99%

Electrophoretically Homogeneous Antibody Synthesized by Spleen Foci of Irradiated Repopulated Mice

Luzzati

Tosi

Carbonara

1970

The Journal of Experimental Medicine

View full text Add to dashboard Cite

106 splenocytes from primed donors were injected, together with sheep erythrocytes (SRBC), into X-irradiated syngeneic mice. 8 days later the spleens were excised and cut into small fragments, keeping track of their location in the organ. Each fragment was cultured individually for 24 hr in the presence of 14C amino acids and the culture fluids were assayed for antibody activity. The antibody-producing fragments were found to be clustered in few restricted areas (foci) surrounded by negative tissue. The anti-SRBC antibody from single foci was purified by absorption on stroma followed by acid elution. Thereafter, it was subjected to electrophoresis and immunoelectrophoresis. The radioautography of the runs showed a considerable degree of homogeneity. Distinct and sharp spikes were localized in the beta and gamma region. The pattern of each focus is unique from the point of view of the number of spikes and their mobility. Eluates obtained from many pooled fragments gave a broad radioactive smear in beta-gamma region. Many foci synthesized antibody migrating as a single band. This homogeneous protein is probably the product of a clone of cells homogeneously differentiated. However, some foci producing two and probably more antibody bands were also encountered. Two interpretations of the finding can be given. Either more than one precursor may participate in the formation of a focus or a differentiation switch may occur during the clonal expansion.

show abstract

Synthesis of antibody by spleen cells after exposure to kiloroentgen doses of ionizing radiation

Cited by 34 publications

References 18 publications

Syngeneic transplantation of hematopoietic stem cells that are genetically modified to express factor VIII in platelets restores hemostasis to hemophilia A mice with preexisting FVIII immunity

Syngeneic transplantation of hematopoietic stem cells that are genetically modified to express factor VIII in platelets restores hemostasis to hemophilia A mice with preexisting FVIII immunity

Antibody Production in Vitro

Electrophoretically Homogeneous Antibody Synthesized by Spleen Foci of Irradiated Repopulated Mice

Contact Info

Product

Resources

About