Immunologic and hematopoietic effects of recombinant human prolactin after syngeneic bone marrow transplantation in mice Rui Sun, Ruth A Gault, Lisbeth.

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Immunologic and hematopoietic effects of recombinant human prolactin after syngeneic bone marrow transplantation in mice Rui Sun, Ruth A Gault, Lisbeth A Welniak, Zhigang G Tian, Susan Richards, William J Murphy Biology of Blood and Marrow Transplantation Volume 9, Issue 7, Pages 426-434 (July 2003) DOI: 10.1016/S1083-8791(03)00107-1

Figure 1 Effects of rhPRL on bone marrow hematopoietic progenitor content after syngeneic bone marrow transplantation. BALB/c mice received 850 cGy and 1 × 106 syngeneic BMC IV. The mice then received PBS or 10 μg rhPRL 3 times per week IP for 3 weeks. Hematopoietic progenitor content was analyzed at various time-points post-SBMT. Progenitor cell CFU-GM (A) and BFU-e (B) assays were performed. Colonies per femur were calculated from the colony frequency and BM cellularity. There were 4 mice for per group at each time point. Data shown are from 3 separate experiments. ∗ = P < .05; ∗∗ = P < .01; ∗∗∗ = P < .001 compared with PBS-treated controls. Biology of Blood and Marrow Transplantation 2003 9, 426-434DOI: (10.1016/S1083-8791(03)00107-1)

Figure 2 Effects of rhPRL on splenic hematopoietic progenitor after syngeneic bone marrow transplantation. BALB/c mice received 850 cGy followed by infusion of 1 million syngeneic BMC IV. The mice then received PBS or 10 μg rhPRL 3 times per week IP for 3 weeks. Mice were euthanized at various time-points post SBMT. Progenitor cell CFU-GM (A) and BFU-e (B) assays were performed. Colonies per spleen were calculated from the colony frequency and splenic cellularity. There were 4 mice for per group at each time point. Data shown are from 3 separate experiments. ∗ = P < .05; ∗∗ = P < .01; ∗∗∗ = P < .001 compared with PBS-treated controls. Biology of Blood and Marrow Transplantation 2003 9, 426-434DOI: (10.1016/S1083-8791(03)00107-1)

Figure 3 Effects of rhPRL on bone marrow B-cell progenitor and mature B-cell content after syngeneic bone marrow transplantation. BALB/c mice received 850 cGy followed by infusion of 1 million syngeneic BMC IV. The mice then received PBS or 10 μg rhPRL 3 times per week IP for 3 weeks. Mice were euthanized at day 21 and day 28. Bone marrow cells were labeled and analyzed with 2-color flow cytometry to determine (A) pre-B cells and (B) B cells. There were 4 mice for per group at each time point. Data shown are from 3 separate experiments. ∗ = P < .05 compared with PBS-treated controls. Biology of Blood and Marrow Transplantation 2003 9, 426-434DOI: (10.1016/S1083-8791(03)00107-1)

Figure 4 Effects of rhPRL on splenic B-cell progenitors and mature B-cell content after syngeneic bone marrow transplantation. BALB/c mice received 850 cGy followed by infusion of 1 million syngeneic BMC IV. The mice then received PBS or 10 μg rhPRL 3 times per week IP for 3 weeks. Mice were euthanized at various time-points and spleen cells were labeled and analyzed with 2-color flow cytometry to determine (A) pre-B cells and (B) B cells. There were 4 mice for per group at each time point. Data shown are from 3 separate experiments. ∗ = P < .05; ∗∗ = P < .01; compared with PBS-treated controls. Biology of Blood and Marrow Transplantation 2003 9, 426-434DOI: (10.1016/S1083-8791(03)00107-1)

Figure 5 Effects of rhPRL on splenic B-cell mitogen responsiveness after congenic bone marrow transplantation. C57BL/6-Ly5.1 mice received 950 cGy total body irradiation followed by infusion of 1 million C57BL/6-Ly5.2 congenic bone marrow cells. The mice then received PBS or 50 μg rhPRL 5 times per week IP for 3 weeks. Mice were euthanized at day 21 and spleen cells proliferative response to LPS was measured as described in “Materials and Methods.” Data shown are from 3 separate experiments. ∗ = P < .05; ∗∗ = P < .01 compared with PBS-treated controls. Biology of Blood and Marrow Transplantation 2003 9, 426-434DOI: (10.1016/S1083-8791(03)00107-1)

Figure 6 Effects of rhPRL on donor engraftment in thymus after congenic bone marrow transplantation. C57BL/6-Ly5.1 mice received 950 cGy total body irradiation. Then received 1 × 106congenic bone marrow cells from C57BL/6-Ly5.2 donor IV. The mice then received PBS or 50μg rhPRL 5 times per week IP for 3 weeks. Thymuses were harvested at various time-points post-CBMT. Donor cell engraftment was calculated from the CD45.1+ cell frequency and cellularity. Mean engraftment and standard error was determined with 4 to 5 mice per group. Data shown are from 3 separate experiments. ∗ = P < .05; ∗∗ = P < .01 compared with PBS-treated controls. Biology of Blood and Marrow Transplantation 2003 9, 426-434DOI: (10.1016/S1083-8791(03)00107-1)

Figure 7 Effects of rhPRL on splenic T-cell subsets after congenic bone marrow transplantation. C57BL/6-Ly5.1 mice received 950 cGy total body irradiation followed by infusion of 1 million C57BL/6-Ly5.2 congenic bone marrow cells. The mice then received PBS or 50 μg rhPRL 5 times per week IP for 3 weeks. Mice were euthanized at day 21 and day 28. Splenocytes were labeled and analyzed with triple-color flow cytometry to determine donor origin and T-cell subsets. There were 4 mice per group at each time point. ∗ = P < .05 compared with PBS-treated controls. Data shown are from 3 separate experiments. Biology of Blood and Marrow Transplantation 2003 9, 426-434DOI: (10.1016/S1083-8791(03)00107-1)

Figure 8 Effects of rhPRL on splenic T-cell mitogen responsiveness after congenic bone marrow transplantation. C57BL/6-Ly5.1 mice received 950 cGy total body irradiation followed by infusion of 1 million C57BL/6-Ly5.2 congenic mice then received PBS or 50 μg rhPRL 5 times per week IP for 3 weeks. Mice were euthanized at day 21 and spleen cells proliferative response to ConA was measured, as described in “Materials and Methods.” Data shown are from 3 separate experiments. ∗P < .05; ∗∗∗P < .001 compared with PBS-treated controls. Biology of Blood and Marrow Transplantation 2003 9, 426-434DOI: (10.1016/S1083-8791(03)00107-1)