Search PubMed⌕ Search

Biomedical subjects

M T Drayson

Publications and source records attributed to M T Drayson.

29 records · Page 2Linked to original sources
PubMed1991

Lymphocyte targeted ricin as a potential therapy for lymphoid malignancy. I. Targeting efficiency.

Lymphocytes were studied as vehicles to target the plant toxin ricin, to lymphoid tissue in rats. Ricin-loaded thoracic duct lymphocyte (TDL) migrated normally into lymph nodes (LN) at 0.5 h, but this process was arrested by 3 h after injection. Ricin was successfully targeted to lymphoid tissue as evidenced by a 4-fold increase in ricin-associated radioactivity in LN, a 10-fold increase in the Peyers patches, a doubling in the spleen and a 35% reduction of radioactivity in the liver compared with free ricin. Nevertheless this represented a considerable shortfall in the expected targeting efficiency. The main problem was found to be high in vivo elution of ricin from TDL (70% within 0.5 h of i.v injection). This and other aspects relevant to maximising targeting efficiency are discussed.

Animals↗
PubMed1989

Fidelity of the repertoire in T cell reconstituted athymic nude rats. Preservation of a deficit in alloresponsiveness over one year.

A single intravenous injection of a relatively small number of T cells contained in the population of rat thoracic duct lymphocytes (TDL) is sufficient to restore to normal the peripheral T cell pool of athymic PVG.rnu/rnu nude rats. The donor T cells expand greater than 10-15-fold, self-renew, and restore immunocompetency to nude recipients permanently (greater than 2 yr). We asked whether the T cell repertoire was affected by the expansion and self-renewal process. Nude recipients were injected with syngeneic PVG TDL that had been allospecifically depleted (negatively selected) by consecutive passage from blood to thoracic duct lymph through two irradiated (DAxPVG)F1 intermediate rats. Negatively selected TDL were tested before transfer by the P----F1 popliteal LN GVH assay and showed a greater than 90% depletion of specific reactivity to DA alloantigens. Surviving cells or their progeny were recovered from LN or TDL of nude recipients 8 and 12 mo after transfer. The deficit in GVH reactivity to the DA haplotype persisted, but normal GVH activity was demonstrated against a third party (AOxPVG)F1 alloantigen. The "hole" in the repertoire could not be attributed to tolerance induced by the co-transfer of contaminating irradiated F1 TDL. PVG TDL passaged consecutively through (AOxPVG)F1 and (DAxPVG)F1 intermediates and devoid of (AOxPVG)F1 cells remained specifically depleted to both AO and DA haplotypes when recovered from nude recipients 4 and 13 mo later, but displayed GVH activity to a third-party (BNxPVG)F1 alloantigen. Thus the exact specificity of the T cell repertoire of the original inoculum was faithfully maintained in nude recipients throughout the initial phase of rapid expansion and the continued self-renewal of the mature peripheral T cell pool.

Animals↗
PubMed1989

The toxicity, distribution and excretion of ricin holotoxin in rats.

The distribution and excretion of the plant toxin ricin were studied in rats after intravenous injection. 125I-labelled ricin was equal in toxicity to native ricin. Following injection, the liver was the major organ of localisation - 46% of injected dose at 0.5 h. The spleen and muscle were next with 9.9% and 13%, respectively, at 0.5 h. Ricin was relatively concentrated in the spleen (33% of injected dose/g of tissue) compared with the liver (7.4%/g) and the bone marrow (5.5%/g). The concentration in the lymph nodes was very low (1.2%/g). Ricin was quickly cleared from the animal; only 11% of the initial radioactivity remained 24 h later with 70% excreted in the urine. Excretion into the intestine via the bile duct was less than 5% by 24 h, 10-12% of the radioactivity was found in the intestinal contents or intestinal wall between 3 h and 12 h, and much of this was reabsorbed since less than 2% was recovered in faeces.

Animals↗
PubMed1989

The origin of T cells in permanently reconstituted old athymic nude rats. Analysis using chromosome or allotype markers.

Athymic PVG-rnu/rnu rats receiving a single intravenous injection of syngeneic euthymic thoracic duct lymphocytes (TDL) develop normal levels of CD4+ T lymphocytes and survive for more than 2 years in a conventional animal house. We investigated the origin of the T cells (and B cells) in reconstituted nude recipients by transferring TDL carrying either the 3T chromosome marker or the RT6b + Igk-1b allotype or the RT7b (leucocyte-common) allotype markers. Karyotype analysis of spleen and lymph node (LN) cells from 1- to 2-year-old PVG-3T/3T-reconstituted nude recipients, stimulated in vitro with phytohaemagglutinin (PHA), unexpectedly revealed that a majority (79-97%) of dividing cells were of nude origin. However, extensive nude cell division was also recorded in PHA-stimulated cultures using mixtures of euthymic (PVG-3T/3T) and unreconstituted nude spleen cells; the assumption that only T cells divide in PHA-stimulated cultures thus appears to be erroneous. In contrast to the karyotype analysis, sIg- RT6b+ LN cells obtained from nude recipients reconstituted 2 years earlier with PVG-RT6b allotype-marked TDL, were all of donor origin with no indication of a nude-derived sIg- RT6a+ population. Igk-1b+ donor B cells were not found in these same recipients. Dual fluorescence analysis of TDL from 18- to 20-month RT7b-reconstituted nudes showed that 91-100% of CD4+ cells were donor-derived. When tested functionally, sIg- RT7b+ (donor) cells, but not sIg- RT7b- (nude-derived) cells, were able to reject skin allografts and induce local graft-versus-host (GVH) responses. Donor T cells, in contrast to CD4+ cells of nude origin, divided extensively in nude recipients; FACS-purified RT7b+ (donor) TDL retransferred from 17-month primary reconstituted nude rats, expanded further (60-100-fold) in secondary nude recipients. In conclusion, only the donor-derived CD4+ cells in reconstituted nude rats displayed T-cell function; evidence to the contrary from karyotype analysis was flawed. At no stage in their life did uninjected or T-cell reconstituted nude rats develop endogenous cells that in any way resembled CD4+ products of the thymus.

Animals↗
PubMed1987

The stable and permanent expansion of functional T lymphocytes in athymic nude rats after a single injection of mature T cells.

Athymic nude rats (PVG.rnu/rnu) were injected at 6 to 10 wk of age with 1 to 200 million thoracic duct lymphocytes (TDL) containing 40 to 60% mature T cells. Thereafter TDL-injected nude recipients were monitored for evidence of T cell function for up to 2 yr. W3/25+ T helper (Th) cells in lymph nodes (LN) increased from 7% at 2 wk to 30% at 8 wk after TDL transfer. The percent of W3/25+ cells remained elevated for the life of the recipient (up to 2 yr), approximating normal levels. The total size of the recirculating pool expanded in TDL-injected nude rats to reach 2/3 the level of euthymic controls by 16 wk, an increase of 10-fold to 15-fold in W3/25+ cells. The expansion of the W3/25+ population was independent of initial TDL dose. With time spleen and LN acquired a normal histological appearance including the development of germinal centres and a marked increase in cellularity in T cell traffic areas. TDL-injected nude rats rejected skin allografts with near normal kinetics. In addition graft vs host (GVH) responsiveness, assessed by the popliteal LN assay, progressively increased reaching a level 9 mo to 1 yr after replacement that resembled the GVH activity in euthymic controls.

Animals↗
PubMed1986

The entry of lymphocytes into stimulated lymph nodes. The site of selection of alloantigen-specific cells.

During a primary response to antigen, lymph nodes (LN) receive antigen-specific lymphocytes (ASL) from the recirculatory pool via specialized postcapillary venules (SPCV). Preliminary experiments demonstrated a three-fold increase in entry of 51Cr-labeled thoracic duct lymphocytes (TDL) into the rat popliteal LN 3-4 days after alloantigenic stimulation. This increased entry was accounted for by an associated 4-5-fold increase in LN blood flow. It was unclear whether recruitment of ASL occurred within the SPCV lumen leading to selective entry of ASL--or, more conventionally, that entry of lymphocytes was a random process and that recruitment of ASL occurred in the extravascular space by selective retention. Thus a double-labeling technique was used to study the migratory characteristics of two populations of thoracic duct lymphocytes (TDL) that had been passaged from blood to lymph. The first population, passaged through a semiallogeneic animal, was negatively selected, and thus 99% depleted of ASL; the second population, passaged through a syngeneic animal, contained a normal complement of naive ASL. The two populations were combined before i.v. injection into syngeneic recipients. At 24 hr the right popliteal LN, which had previously been stimulated with the specific antigen, showed a preferential localization of the TDL containing ASL compared with the left (unstimulated) popliteal LN. This preferential localization was not present at 1 hr after injection. In the light of background data on the kinetics of lymphocyte traffic it is concluded that ASL were selected not at the luminal surface of SPCV but in the LN after they had left the bloodstream.

Animals↗
PubMed1985

The effects of cyclosporin on lymphocyte activation in a systemic graft-vs.-host reaction.

We have investigated the effects of cyclosporin (CsA) on each of three stages of lymphocyte activation in vivo viz. sequestration of alloantigen-reactive lymphocytes from the circulation into the spleen and lymph nodes, blast transformation and induction of DNA synthesis in the activated cells and release of these cells and their progeny into the circulation. Parental strain lymphocytes injected i.v. into semi-allogeneic rats and recovered from the thoracic duct within 36 h are profoundly unresponsive in a local graft-vs.-host assay to the alloantigens of the F1 hybrid but have normal activity against unrelated alloantigens (negative selection). CsA treatment of the F1 hybrid recipients did not prevent this selective sequestration of antigen-reactive cells. In the untreated F1 hybrid, from 36 h after injection, large numbers of dividing blast cells were released into the lymph. These cells did not appear in the lymph of recipients treated with CsA. However, CsA did not prevent the activation of cells sequestered in the spleen or lymph nodes as assessed by [3H] thymidine incorporation and autoradiography. This unexpected finding suggests that CsA inhibits lymphocyte responses to alloantigens in vivo after DNA synthesis which is a later stage than the in vitro studies have shown.

Animals↗
PubMed1984

Afferent lymph and lymph borne cells: their influence on lymph node function.

In AO rats the afferent lymphatics to the right cervical lymph nodes (LN) were interrupted and the LN were encased in silicone rubber tubes to prevent reunion of the lymphatics. At regular intervals over the next 12 weeks the following were measured in comparison with the intact contralateral LN - LN weight, influx of lymphocytes from the blood, blood flow, the incorporation of 125IUdR and the incorporation of 35S-sulphate into high endothelial venules (HEV). Systematic histological observations are also reported. One day after deafferentization lymphocyte influx was significantly reduced although blood flow was unchanged and a temporary increase in LN weight was associated with crowding of the lymphatic sinuses with small lymphocytes. The subsequent decline in lymphocyte influx was biphasic and quicker than the decline of other parameters--being undetectable by 6 weeks. Flattening of HEV and diminished secretion of 35S-sulphate was noted at 1 week and progressive degeneration and eventual disappearance of the HEV network was seen by 6-12 weeks. Doubtlessly because of lack of antigenic stimulation 125IUdR incorporation, and numbers of lymphoblasts, plasma cells and finally germinal centres were progressively reduced. The numbers of macrophages and interdigitating cells (IDC) were greatly reduced by 3 weeks and very few were present at 6 weeks probably because most or all arrive in afferent lymph and have a limited life span in the LN. At 12 weeks the LN was difficult to recognize as such since only stromal cells and occasional small lymphocytes remained. In supplementary experiments u.v. irradiation of the LN at the time of deafferentization reduced lymphocyte influx without affecting blood flow suggesting that a u.v. sensitive cell like the IDC may influence lymphocyte influx. In conclusion the involution of the deafferentized LN is partly due to the lack of antigen but progression to the complete loss of specialized structure and function is probably due to lack of other factors including non-lymphoid cells that normally arrive in afferent lymph.

Animals↗
PubMed1981

The sequence of changes in blood flow and lymphocyte influx to stimulated rat lymph nodes.

The rat popliteal lymph node was studied from 1 hr to 8 days after the footpad injection of either sheep erythrocytes or syngeneic rat erythrocytes. The following were measured relative to the contralateral (unstimulated) lymph node: (i) blood flow; (ii) lymph node weight; (iii) influx of lymphocytes from the blood; (iv) [3H]-thymidine incorporation; (v) [35S]-sulphate incorporation into macromolecular form (chiefly by high endothelial venules). After the arrival of sheep erythrocytes all five quantities showed substantial increases which began in a definite sequence. The blood flow started to rise first and may have been the main factor contributing to the later increase in lymphocyte influx. Increased sulphate incorporation began later than the rise in lymphocyte influx. After the injection of rat erythrocytes a small increase in lymphocyte influx was found without a corresponding increase in blood flow. In rats irradiated before the footpad injections lymphocyte influx increased three-fold after sheep erythrocytes, rat erythrocytes or PBS, again without a corresponding increase in blood flow. Thus while variation in blood flow to high endothelial venules is one important factor in determining the supply of lymphocytes to the lymph node other factors are operative in certain situations.

Animals↗