Figure 5
Figure 5. ITI of cells or drugs can rejuvenate and enhance thymopoiesis of aged recipients. (A) Eleven-month-old (old) or 5-week-old (young) C57BL/6 recipients received 6000 C57BL/6.CD45.1+ LSK cells via ITI 2 hours after sublethal TBI. Thymuses and spleens were harvested on day 60 after radiation and analyzed for donor and host origin and CD3+ populations. Mean and SEM of combined data from 2 independent experiments are presented (n = 5). (B) Old and young C57BL/6 recipients received ITIs with syngeneic LSK cells as described in panel A. Splenocytes were analyzed for B cells, natural killer (NK) cells, and myeloid cells of donor origin on day 60 after radiation. Mean and SEM of combined data from 2 independent experiments are presented (n = 5). (C) Twelve-month-old C57BL/6 mice received 2.5 μg KGF or 500 ng IL-7 or 500 ng IL-22 via ITI. Age-matched control mice received no injection. No irradiation was given to any of the recipients. Thymuses were harvested on day 30 after injection and analyzed for total thymocyte numbers. Mean and SEM of 1 of 3 independent experiments are presented (n = 4). (D) Six-week-old C57BL/6 mice received 2.5 μg KGF or 500 ng IL-7 or 500 ng IL-22 via ITI 24 hours prior to sublethal TBI. Age-matched control mice received no injection. Thymuses were harvested on day 30 after irradiation and analyzed for total thymocyte numbers. Mean and SEM of 1 of 2 independent experiments are presented (n = 5). (E) Nine-month-old BALB/c mice were intrathymically injected with either PBS or 3 different doses of IL-7 cytokine: 20 ng, 100 ng, or 500 ng. Sublethal irradiation was performed on day 1 postinjection. Thymuses were harvested on day 30. Mean and SEM are presented (n = 4-5). (F) Nine-month-old BALB/c mice received a sublethal dose of TBI and a single ITI with either 2.0 μg of KGF on day −28 or 500 ng of IL-7 on day −1 or both. Age-matched control mice received no injection. Thymuses were harvested on day 30 after irradiation and analyzed for total CD45+ thymocyte numbers. Mean and SEM of 1 of 2 independent experiments are presented (n = 4-6). (G) Nine-month-old BALB/c mice were treated as described in panel F. Splenocytes were harvested on day 30 after irradiation and analyzed for total CD3+ T cells. Mean and SEM of 1 of 2 independent experiments are presented (n = 4-6). XRT, radiation therapy.

ITI of cells or drugs can rejuvenate and enhance thymopoiesis of aged recipients. (A) Eleven-month-old (old) or 5-week-old (young) C57BL/6 recipients received 6000 C57BL/6.CD45.1+ LSK cells via ITI 2 hours after sublethal TBI. Thymuses and spleens were harvested on day 60 after radiation and analyzed for donor and host origin and CD3+ populations. Mean and SEM of combined data from 2 independent experiments are presented (n = 5). (B) Old and young C57BL/6 recipients received ITIs with syngeneic LSK cells as described in panel A. Splenocytes were analyzed for B cells, natural killer (NK) cells, and myeloid cells of donor origin on day 60 after radiation. Mean and SEM of combined data from 2 independent experiments are presented (n = 5). (C) Twelve-month-old C57BL/6 mice received 2.5 μg KGF or 500 ng IL-7 or 500 ng IL-22 via ITI. Age-matched control mice received no injection. No irradiation was given to any of the recipients. Thymuses were harvested on day 30 after injection and analyzed for total thymocyte numbers. Mean and SEM of 1 of 3 independent experiments are presented (n = 4). (D) Six-week-old C57BL/6 mice received 2.5 μg KGF or 500 ng IL-7 or 500 ng IL-22 via ITI 24 hours prior to sublethal TBI. Age-matched control mice received no injection. Thymuses were harvested on day 30 after irradiation and analyzed for total thymocyte numbers. Mean and SEM of 1 of 2 independent experiments are presented (n = 5). (E) Nine-month-old BALB/c mice were intrathymically injected with either PBS or 3 different doses of IL-7 cytokine: 20 ng, 100 ng, or 500 ng. Sublethal irradiation was performed on day 1 postinjection. Thymuses were harvested on day 30. Mean and SEM are presented (n = 4-5). (F) Nine-month-old BALB/c mice received a sublethal dose of TBI and a single ITI with either 2.0 μg of KGF on day −28 or 500 ng of IL-7 on day −1 or both. Age-matched control mice received no injection. Thymuses were harvested on day 30 after irradiation and analyzed for total CD45+ thymocyte numbers. Mean and SEM of 1 of 2 independent experiments are presented (n = 4-6). (G) Nine-month-old BALB/c mice were treated as described in panel F. Splenocytes were harvested on day 30 after irradiation and analyzed for total CD3+ T cells. Mean and SEM of 1 of 2 independent experiments are presented (n = 4-6). XRT, radiation therapy.

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