Mice of several inbred strains were studied for responses to Treponema pallidum (Nichols). Mice were injected with viable treponemes and observed daily for the presence of lesions or signs of illness. Although none of the mice developed detectable lesions, they did develop specific anti-T. pallidum antibodies that were observed as early as 21 days after inoculation with viable T. pallidum. The antibodies were detected by an enzyme-linked immunosorbent assay and by passive haemagglutination.
We have previously reported that platelet concentrates (PC) may be irradiated with ultraviolet light (UVL) in a cryostorage pack such that mixed lymphocyte reactions (MLR) are abolished whilst satisfactory platelet function is retained during subsequent storage using Fenwal PL-1240 containers. We have now studied both platelet structure and function after irradiation in DuPont Stericell bags which are both UV-permeable and biocompatible. The irradiation dosage was 3000 Joules/m2 of UVL at a mean wavelength of 310 nm; a dose previously shown to abolish MLR. No detriment to platelet function was observed when compared to control as measured by aggregation responses to adenosine diphosphate (ADP), collagen and ristocetin, hypotonic shock response and pH during 5 d of storage. Lactate levels were significantly higher (P less than 0.01) and glucose levels lower (P less than 0.01) in UV-treated PC, although in the majority the lactate level did not exceed 20 mmol/l. Betathromboglobulin and platelet factor 4 levels were higher during storage in the UV group, the latter reaching significance (P less than 0.05). When whole platelets and platelet membranes were stained with Coomassie blue or Periodic Acid-Schiff's reagent after electrophoresis in polyacrylamide gels no obvious alterations to major membrane constituents were observed on days 1 and 5 of storage. Paired in vivo autologous studies in healthy volunteers using 111Indium-labelling were performed at the end of 5 d of storage. The UV-treated platelets gave satisfactory and equivalent results for recovery, half life and survival when compared to controls. We conclude that PC irradiated with UVL and stored for 5 d in DuPont Stericell containers appear suitable for transfusion and may prove to be nonimmunogenic. Further in vivo studies of haemostatic efficacy and recipient alloimmunization are now warranted.
Passenger lymphocytes in platelet concentrates (PCs) may induce the formation of lymphocytotoxic antibodies (LCTAbs) and subsequent refractoriness to platelet transfusions. Ultraviolet (UV) irradiation can prevent lymphocytes' acting as stimulator or responder cells in mixed-lymphocyte reactions (MLRs) and could theoretically prevent LCTAb formation in vivo. A system has been devised for the delivery of UV irradiation to PCs; platelet storage characteristics and MLRs were evaluated in UV-irradiated PCs harvested from healthy donors with the Haemonetics V50 and PCS cell separators. MLR and response to phytohemagglutinin stimulation were abolished by a dose of 3000 joules per m2 at a mean wavelength of 310 nm. Platelet aggregatory responses to adenosine diphosphate (ADP), ristocetin, collagen and epinephrine, hypotonic shock response, and pH showed no important differences when control PCs and PCs irradiated as above were compared during 5 days of storage in Fenwal PL-1240 packs. Lactate production during storage was significantly higher in UV-treated PCs (p less than 0.001), but values did not exceed 20 mmol per L. UV transmission at 310 nm in standard blood product containers, including the Fenwal PL-146, PL-1240, and PL-732, was low (less than 30%), but it was acceptable in the Delmed Cryostorage and DuPont SteriCell packs (greater than 50%). UV irradiation may provide a simple and inexpensive means of producing nonimmunogenic PCs.
To investigate the development of the humoral immune response of mice to infection with Treponema pallidum, Balb/c and C57Bl mice were injected intradermally with 10 x 10(6) virulent organisms. Serum samples were taken from the mice at weekly intervals after infection and used in the electrophoretic transblotting technique to detect T pallidum and T phagedenis biotype Reiter antigens. The serum samples taken from infected mice at 21, 42, 84, and 126 days after infection recognised two to 15 T pallidum antigens, with a gradual but continual increase in the number of antigens recognised. The same antisera to T pallidum recognised five cross reactive T phagedenis biotype Reiter antigens. Mice injected with 10 x 10(6) heat killed T pallidum organisms failed to recognise T pallidum antigens, as shown by the blotting technique. When mice infected with T pallidum were given antibiotics, the development of the humoral response was interrupted. These experiments indicated that mice respond more slowly than rabbits to T pallidum, which may be because T pallidum is weakly immunogenic in mice.
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