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Biomedical subjects

W R Clark

Publications and source records attributed to W R Clark.

At least 73 records · Page 4Linked to original sources
PubMed1991

Lack of target cell participation in cytotoxic T lymphocyte-mediated lysis.

Data on the subject of cell-mediated cytotoxicity suggest that no single mechanism is likely to provide a satisfactory explanation of this process. Lytic pathways have been proposed that involve both the effector cell and the target cell as active participants. In this report we describe a system in which the target cell is rendered unable to participate in its own demise. Using sheep E derivatized with CD3 antibodies, we show that metabolic inhibition of SRBC by depleting intracellular ATP with iodoacetamide, or even conversion of SRBC to "ghosts" by hypotonic lysis and resealing, has no effect on cytolysis. In the presence of EGTA or cholera toxin, both of which inhibit CTL degranulation, there is a strong suppression of both serine esterase release and cytolysis. These data show clearly that in some situations CTL are able to lyse target cells without any active participation by the target cells themselves.

Adenosine Triphosphate↗
PubMed1991

Methylprednisolone does not protect the lung from inhalation injury.

Most clinical studies suggest that corticosteroids are contraindicated in the treatment of acute smoke inhalation. However, they are still used in critical situations with the hope that they might reverse the acute pathophysiological responses to smoke inhalation and thus reduce the severity of the illness or make survival possible. These experiments were done to study the effect of methylprednisolone on the response to smoke inhalation in anaesthetized mongrel dogs. Three experimental protocols were followed: (I) haemodynamics, gas exchange, lung compliance, and lung water were evaluated; (II) pulmonary vascular permeability was assessed by cannulating the afferent tracheobronchial lymphatic and calculating the osmotic reflection coefficient (sigma d) at high lung lymph flows; (III) pulmonary surfactant function was studied using a Wilhelmy balance. Methylprednisolone alone did not alter any measured values compared with those seen in control animals. Treatment with methylprednisolone (30 mg/kg) prior to smoke exposure did not attenuate any of the adverse responses typically seen after smoke inhalation. These data indicate that methylprednisolone does not protect the lung from the acute physiological consequences of inhalation injury.

Animals↗
PubMed1991

Cardiac transplantation in patients with preexisting malignancies.

A history of preexisting malignancy has been considered a contraindication to cardiac transplantation. The reasons for this prejudice include concerns about potentially deficient intrinsic immunomodulation and fear of cancer recurrence (or development of second cancers) because of therapeutic immunosuppression. In the past four years at the Northern Indiana Heart Institute seven patients with preexisting malignancies underwent cardiac transplantation. Their two-year survival rate was 100%, which is comparable to a rate of 81% in non-malignancy patients. After an average 31 months of follow-up (range = 6-56 months), only one patient has had a recurrent tumor (basal cell carcinoma). Statistical comparison of immunosuppression dosages, incidences of rejection, and incidences of infections between patients with preexisting malignancy and those without preexisting malignancy was performed. We found that the only significant difference was an increased number of infections in preexisting malignancy patients. Additionally, we found no difference in the incidence of posttransplant coronary artery disease in the preexisting malignancy group when compared with those patients without preexisting malignancies. This study demonstrates that patients who have been successfully treated for malignancies have no greater incidence of rejection than those patients without preexisting malignancy. Furthermore, preexisting malignancy patients require no significant modulation of immunosuppression. Although preexisting malignancy patients have a higher incidence of infections than patients without preexisting malignancy, their two-year survival is not worse than the patients without preexisting malignancy.

Adult↗
PubMed1990

Expression of mRNAs for pore-forming protein and two serine esterases in murine primary and cloned effector lymphocytes.

The cDNAs encoding several proteins present in the granules of cytolytic effector lymphocytes have now been cloned. These include the cytolytic pore-forming protein (PFP) or perforin, and at least six serine esterases (SE), also called granzymes. The cDNA probes for PFP, SE-1, and SE-2 are used here to study the expression of these proteins in murine primary effector lymphocytes. Among the stimuli effective in inducing the expression of PFP, SE-1, and SE-2 were recombinant interleukin-2, the lectin concanavalin A in the presence of phorbol esters, and allogeneic cells in mixed lymphocyte cultures. Some correlation was seen between the levels of PFP and SE mRNAs and cytotoxicity measured in a standard 51Cr release assay. We also examined a panel of 13 cloned cytotoxic T lymphocyte (CTL) lines and found that mRNAs for PFP and SE-2 were expressed in all CTL lines, including some that were previously considered not to produce PFP. Twelve of the 13 CTL lines also proved to possess the mRNA for SE-1. One thymoma cell line, TIMI.4, did not express mRNA for PFP, although it expressed mRNA for SE-1 and SE-2.

Animals↗
PubMed1990

Distribution of extravascular lung water after acute smoke inhalation.

Anesthetized dogs with thoracotomy were injected with Evans blue dye and were exposed acutely (5 min) to wood smoke inhalation. Thin slices from freeze-dried samples were photographed and assessed for periarterial and perivenous cuff area and for blue coloration with a score of 0 to 5. Bloodless extravascular lung water (EVLW) was also measured. The smoke-exposed animals were compared with controls and with animals exposed to alloxan or to high-pressure-induced pulmonary edema. EVLW at 2 h after smoke (6.46 +/- 0.80) was above control value (4.30 +/- 0.63) but not different from the alloxan (6.13 +/- 0.70) or high-pressure (6.88 +/- 1.30) groups. Despite the similarity in EVLW in the edematous lungs, there were marked differences in the intensity of blue color and size of cuffing around arteries and veins: the smoke, alloxan, and high-pressure groups had blue color scores of 1.0 +/- 0.1, 2.9 +/- 0.3, and 0.3 +/- 0.1, respectively. These scores indicated a large increase in microvascular permeability to proteins in the alloxan group, a moderate increase in the smoke group, and minimal change in the high-pressure group. The perivascular cuff area was largest in the alloxan group and moderate in the smoke and high-pressure groups. The cuff area was higher for arteries than for veins in all groups except the 0.5-h smoke group. We conclude that smoke inhalation causes a moderate increase in permeability and EVLW compared with alloxan. The extravascular lung water accumulates preferentially around the arteries, but the size of the perivascular cuff is not similar for all causes of pulmonary edema.

Alloxan↗
PubMed1989

Evidence for multiple lytic pathways used by cytotoxic T lymphocytes.

Previous data generated by ourselves and others questioned the role of degranulation as a mechanism to explain CTL-mediated cytotoxicity. In this report we examine this tissue in greater depth. CTL-mediated lysis was probed with three different inhibitors. 4,4'-diisothiocyano-2,2'-disulfonic acid stilbene inhibits degranulation in a wide range of cell types, including CTL. EGTA, through chelation of Ca2+, also inhibits degranulation processes in CTL, and would inhibit other events or processes dependent on extracellular Ca2+. We also used prolonged exposure to PMA to exhaust PKC activity in CTL. Using these inhibitors, we have defined three pathways of lysis used by CTL. One pathway requires Ca2+, is PMA sensitive, but does not depend on degranulation. The second pathway is independent of Ca2+, is not PMA sensitive, and also does not depend on degranulation. All primary CTL and cloned CTL lyse most target cells via pathway I. However, when confronted with certain target cells (which we have referred to previously as Ca2+-independent target cells), pathway II is induced. When pathway II is induced, pathway I apparently shuts down. We show here that pathway II does not depend on protein synthesis, and that it also leads to DNA solubilization in target cells. A limited number of cloned CTL use pathways I and II as just described, but use in addition, and simultaneously, a third pathway that appears to involve degranulation. This pathway is seen irregularly in most CTL clones, and may be influenced by levels of IL-2 in the culture medium.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed1989

Alcohol exposure in utero results in diminished T-cell function and alterations in brain corticotropin-releasing factor and ACTH content.

The long-term teratogenic effects of prenatal ethanol exposure during the last week of gestation on immune responsiveness and levels of pituitary ACTH and hypothalamic corticotropin-releasing factor (CRF) were examined in Sprague-Dawley rats. Immune responsiveness was measured by T-lymphocyte proliferation in response to mitogenic stimulation with Con A (3 micrograms/ml) in spleen and thymus cells of 21-old-day male rats who were exposed to alcohol in utero. The proliferative response was 8-fold lower in spleen and twofold lower in thymus cells from alcohol-exposed animals compared to responses measured in control rats. Thymus weight was significantly smaller at birth in alcohol exposed males, but significantly larger at 21 days of age compared to controls. Alterations in the content of ACTH and CRF, hormones, known to be direct or indirect modulators of immune responsiveness, were also observed in alcohol exposed males. Hypothalamic content of CRF and pituitary content of ACTH were significantly lower in alcohol exposed males on postnatal Day 1, but hypothalamic ACTH content was significantly higher compared to controls. These results indicate that alcohol exposure during the last week of gestation can produce alterations of the hypothalamic-pituitary-adrenal function in addition to teratogenic effects on the immune system which have been previously observed only with a much longer alcohol exposure regimen.

Adrenocorticotropic Hormone↗
PubMed1989

Wood smoke inhalation increases pulmonary microvascular permeability.

The effect of wood smoke inhalation (SI) on pulmonary vascular permeability was studied in open-chested, anesthetized dogs. Animals were divided into two groups. A prenodal lymphatic vessel was cannulated in group I (n = 7), and baseline (BL) lung lymph flow (QL) and lymph (CL) and plasma (CP) protein concentrations were measured. The animals' lungs were then ventilated with wood smoke for 5 minutes. Left atrial pressure (Pla) was increased above baseline (mean 16.7 +/- 2.2 mm Hg), and the ratio of CL to CP was used to assess endothelial permeability at high lymph flows. There was little change in either QL (BL: 27 +/- 9; SI: 27 +/- 5 microliters/min) or CL/CP (BL: 0.76 +/- 0.03; SI: 0.74 +/- 0.02) after SI at normal Pla. Elevation of Pla caused a significant increase in QL (136 +/- 15 microliters/min), but CL/CP (0.67 +/- 0.02) failed to decrease significantly at high lymph flows. In group II (n = 15) total protein concentration of airway fluid was compared with that of plasma after smoke inhalation, intravenous alloxan, and increased Pla. The ratio of protein concentration in airway fluid to plasma after SI (0.70 +/- 0.07) was greater than that obtained with increased Pla (0.64 +/- 0.07) but less than that after alloxan (0.85 +/- 0.04). These data indicate that SI in the dog results in a moderate increase in pulmonary vascular permeability that is less severe than that induced by alloxan.

Animals↗
PubMed1988

Renal adenoma in a cat.

Renal adenoma, a benign tumor of epithelial origin, was diagnosed in a 10-year-old cat. Clinical signs included decreased appetite and weight loss. Nephrectomy resulted in clinical improvement. Seven months after surgery, the cat died, and pulmonary papillary adenocarcinoma was discovered. The pulmonary neoplasm differed markedly from the renal neoplasm.

Adenocarcinoma, Papillary↗
PubMed1988

Cloned cytotoxic T lymphocytes as target cells. II. Polarity of lysis revisited.

The original polarity of lysis experiments suggested that CTL are themselves sensitive to whatever mechanism it is that CTL use to lyse their targets. This concept has placed certain limitations on possible mechanisms of lysis by CTL. Recently, we found in studies with cloned CTL as targets that cloned CTL are in fact highly resistant to lysis by other CTL, as well as to their cytotoxic granule proteins. We show here that although cloned CTL are extremely resistant to lysis by primary and cloned CTL, they are readily inactivated functionally by all primary CTL and by at least one CTL clone. Moreover, cloned CTL are also functionally inactivated by cytotoxic granule proteins. The activation of CTL, which we call inhibitin, is Ca2+ insensitive and distinct from hemolytic activity, and is, thus, unlikely to be perforin. These experiments suggest a possible alternative interpretation of the original polarity of lysis experiments.

Animals↗
PubMed1988

The role of accessory molecules in T-helper activation induced by antigen, lectin, or CD3 antibodies.

We examined the role of L3T4 and LFA-1 molecules in T-helper-cell activation, under conditions where the physical stability of T helper-accessory cell interactions was not an issue. T-helper hybridomas were activated by accessory cells coated either with concanavalin A (Con A) or with CD3 antibodies. Activation of the T helper cells was measured by microtubule-organizing center (MTOC) reorientation as an early activation event, and by interleukin-2 (IL-2) production as an indication of a fully matured response. Both parameters were strongly blocked by L3T4 and LFA-1 antibodies in the case of Con A activation. In the case of stimulation with accessory cell-bound CD3 antibody, activation was blocked by LFA-1 but not L3T4 antibody. These results support the notion that L3T4 and LFA-1 molecules play more than a simple adhesion role in T-cell activation. The differential effect of L3T4 antibody in the case of Con A activation vs CD3 activation is consistent with the possibility that L3T4 and the alpha/beta portions of the T-cell receptor must interact during antigen- and lectin-stimulated T-cell activation.

Animals↗
PubMed1988

Cloned cytotoxic T lymphocyte target cells fail to induce early activation events in effector cytotoxic T lymphocytes.

We have explored further the basis for resistance of cloned cytotoxic T lymphocytes (CTLs) to cell-mediated cytotoxicity. We find that most cloned CTLs recognized as specific target cells by other cloned CTLs used as effector cells fail to activate three early events that may be critical in triggering lysis in the effector CTLs: Ca2+ influx, microtubule organizing center (MTOC) reorientation, and serine esterase release. To the extent that any or all of these events are involved in activation or expression of the lytic pathway in effector CTLs, our results suggest that in addition to being inherently resistant to cytotoxic granule extracts, many CTLs are also unable to induce lytic function in other (effector) CTLs. We have found one CTL clone that can respond to recognizable cloned CTL target cells with at least MTOC reorientation and serine esterase release, although the target CTLs are still not lysed. In this case, the resistance of the target CTL to lysis may be due solely to its resistance to cytoplasmic granule contents.

Animals↗
PubMed1988

The toxic shock syndrome in a burn victim.

A case of toxic shock syndrome (TSS) in a 6-year-old boy with a 5 per cent body surface area partial thickness scald burn is reported. Biobrane was the wound dressing used initially. The clinical and laboratory characteristics of the syndrome are listed. The syndrome is caused by an exotoxin of Staph. aureus and can occur in the absence of invasive infection. The syndrome, first described in menstruating women who used tampons, has occurred in patients with a variety of wounds. Reports of the TSS syndrome in burn victims are limited to children whose wounds were covered. Children are most vulnerable to the development of TSS because of their limited prior exposure to the toxin.

Biocompatible Materials↗
PubMed1988

Smoke inhalation.

An attempt has been made to review the characteristics of fire and smoke and the epidemiology of smoke inhalation to identify some of the many variables which interact to control the severity of the injury. An experimental model appropriate to study the pulmonary injury of smoke victims who survive to enter the health care system is described. Experiments which define how smoke damages the lung are reviewed in an effort to explain why the smoke-damaged lung is vulnerable to additional stress and why those with an injured lung and a burn have such a high mortality rate.

Adult↗
PubMed1988

Effects of crystalloid on lung fluid balance after smoke inhalation.

Inhalation injury occurs in 21% of flame burn victims who require large fluid volumes for resuscitation and have a mortality rate greater than 30%. This study was done to determine how vulnerable the smoke-injured lung is to fluid accumulation when crystalloids are infused rapidly. Mongrel dogs were exposed to smoke and 10% body-weight Ringer's lactate in three groups: (I) fluid only, (II) smoke only, and (III) smoke and fluid. The increase in wet-dry lung weight ratio was 2% in Group I, 28% in Group II, and 42% in Group III, consistent with pulmonary edema present only in Group III. The decrease in colloid oncotic pressure was similar in both of the groups that were given fluid, and the rise in the surface tension minimum of lung extracts was similar in both of the groups that were exposed to smoke. The smoke-injured lung loses the ability to protect itself when challenged with fluid. Reduced oncotic pressure is not responsible. Changes in microvascular pressure, endothelial and epithelial damage, and surfactant inactivation interact to cause this increase in extravascular lung water.

Animals↗