Clonal character of F1 hybrid lymphocyte subset recognition of parental cells in one-way mixed lymphocyte cultures.

Proliferation of F(1) hybrid lymphocytes in mixed lymphocyte cultures is stimulated by mitomycin-blocked parental cells. The demonstration of this phenomenon using F(1) hybrids derived from congenic lines of mice establishes that the stimulation is controlled by genes in or closely linked to the major histocompatibility locus chromosome region. In agreement with the finding that tumor-bearing mice have an increased capacity for primary alloantigen recognition, it was observed that the F(1) hybrid response to parent was also augmented by tumor bearing. Chromosomal analysis of dividing cells in one-way mixed cultures confirms that F(1) cells, and not the blocked parental cells, enter mitosis. Stimulation of F(1) cells by a soluble mediator liberated by the parental cells was not observed and mitomycin blocking of parental cells seems to be a completely effective blocking agent ensuring that parental cells can not enter DNA synthesis. The specificity and clonal nature of F(1) recognition of parent was demonstrated using a 5-bromodeoxyuridine-suicide procedure. Distinct clones of lymphocytes in F(1) spleen cell populations seem to recognize one or the other parent, but not both, in such experiments. These observations and others in tumor systems suggest that most or all heterozygous organisms may possess potentially self-reactive clones of lymphocytes.

Regulation of B-cell proliferative responses to lipopolysaccharide by a subclass of thymus T cells.

When thymus cells which are unresponsive to LPS are combined with numbers of peripheral lymphoid cells giving minimal responses to LPS, synergistic incorporation of [3H]thymidine occurs. Synergy requires that both components proliferate, but most of the augmented response is the result of peripheral cell proliferation. The thymus cell is a T cell of variable density, low in thy-1.2 antigen, not concanavalin A responsive, present in the major thymus subpopulation, and may be from lipopolysaccharide (LPS)-unresponsive strains. The peripheral cell is sensitive to anti-IgG or IgM plus complement (C'), resistant to anti-Thy-1.2 and C', exhibits adherence properties of B lymphocytes, and must be from LPS-responsive strains. Synergistic responses depend on critical thymus/peripheral cell ratios, inhibition occurring at high peripheral cell numbers. The data provide evidence that B-cell proliferative responses to LPS may be regulated by a subclass of thymus T cells.

Generation of cytotoxic effector cells by immunocompetent thymus cell subpopulations.

Immunocompetent mouse thymus cell subsets (TH-2) cultured with allogeneic TH-2 cells, required a peripheral lymphoid cell in order to generate specifically cytotoxic T cells (CTL) in vitro. The helper cell is identified as a B cell, and can be supplied either in the mixture of target or responding peripheral cells, or in optimal ratios of 10:1 to 20:1 as nonproliferating cells syngeneic to the responder, when both responder and target are TH-2. Allogeneic helper cells are much less effective. Kinetic studies indicate that helper cell activity involves initial interaction with stimulator cell and is then required throughout the period of generation of cytotoxic lymphocytes. Addition of critical levels of certain IgG preparations, particularly IgG2A appears to support generation of CTL in TH-2/TH-2 cultures but is considerably less effective than intact B cells. Cytotoxicity generated under these conditions has the same genetic requirements for I-region histoincompatibility between stimulator and responder, and for K or D identity of stimulator and target cell, as has been reported for peripheral cell CTL.

Immunologic properties of mouse thymus cells. Identification of T cell functions within a minor, low-density subpopulation.

The functional attributes of minor subpopulations of mouse thymus cells derived by bovine serum albumin density gradient centrifugation, cortisone treatment, or selective depletion by anti-TL or anti-theta treatment have been examined. A subpopulation derived in each fashion contains the cells required to evoke graft-versus-host reactions in neonatal F(1) hybrid recipients and to be stimulated by alloantigens in vitro in one-way mixed lymphocyte cultures and by phytohemagglutinin. The functions of this subpopulation are abrogated by treatment with anti-H-2 plus complement and by high concentrations of anti-theta. A tentative ordering of the various thymus cell subpopulations, on the basis of these and other data, is described.

Cellular recognition in vitro by mouse lymphocytes. Effects of neonatal thymectomy and thymus graft restoration on alloantigen and PHA stimulation of whole and gradient-separated subpopulations of spleen cells.

The effects of thymectomy and thymus graft restoration upon the in vitro primary responses to alloantigens and PHA have been studied. It has been found that neonatal thymectomy substantially eliminates both PHA reactivity and responsiveness to alloantigens assayed in vitro in host spleen cell populations. Analysis of albumin density gradient-separated subpopulations of the spleen and thymus in such animals was also performed. It was found that the total and proportional representation of the individual density subpopulations was identical in neonatally thymectomized, in normal, and in thymectomized and thymus graft-restored animals. Therefore, thymectomized mice appear to retain a nonfunctioning, small, dense, lymphocyte population. Reconstitution of thymic-dependent in vitro reactivity was nearly complete when syngeneic, but not allogeneic or semisyngeneic thymus was employed. Occasional partial restoration did occur when F(1) thymus was employed, but never when allogeneic thymus was grafted. The grafted thymus contained PHA and alloantigen-reactive cells in a large, less dense B layer subpopulation, whereas the restored animals, as in the case of normals, showed these reactivities to be a property of a small, more dense cell population.

Tumor-secific immunity to chemically induced tumors. Evidence for immunologic specificity and shared antigenicity in lymphocyte responses to soluble tumor antigens.

Experiments were designed to explore the apparent paradox that methylcholanthrene-induced tumors of mice evoke tumor-unique transplantation immunity but reveal almost complete cross-reacting antigenicity in tests of lymphocyte behavior in vitro. The approach involved use of tumor membranes solubilized in 3 M KCl, employed both as the stimulating antigen source in a new in vitro proliferation assay of lymphocyte recognition, and as immunogens in vivo. The kinetics of the assay resembled those of in vitro tests of mitogen or specific antigen stimulation in other systems. Lymphoid cell proliferation was assessed in peripheral blood leukocytes, lymph nodes (LN), and spleen over the course of tumor bearing, and in animals immunized by tumor amputation or with the solubilized antigens. The pattern of spread of reactivity was from regional LN to spleen, peripheral blood, and nonregional nodes in each circumstance. An unexplained low antigen dose inhibitory phenomenon was encountered in spontaneously proliferating cell subpopulations taken from some tumor-bearing animals. In vitro responses to some but not all solubilized antigens made from multiple syngeneic tumors were detected in each circumstance. The soluble antigens also induced shared resistance to some tumors. The patterns of spread of responsiveness to syngeneic tumor antigens, the time-course, and relative intensity were most compatible with independent clonal responses to multiple tumor-borne antigens, some but not all of which are shared in any family of syngeneic tumors.

Evidence for a B-cell -like helper function in mixed lymphocyte culture between immunocompetent thymus cells.

The immunocompetent subpopulation by mouse thymus cell (TH-2) was isolated by buoyant density centrifugation and by hydrocortisone pretreatment. TH-2 cells undergo a proliferative one-way or two-way mixed lymphocyte culture (MLC) response only when cultured with allogeneic or congenic peripheral lymphoid cells. However, mixtures of allogeneic TH-2 cells alone do not proliferate in either one-way or two-way MLC reactions. Such MLC mixtures are proliferative only if mitomycin-blocked peripheral lymphoid cells are also present in the mixture. The peripheral helper cell has been found to be of low net density, non-adherent, insensitive to anti-thy-1 serum cytotoxicity, but sensitive to the cytotoxic effets of anti-immunoglobulin serume plus complement. The helper effect does not depend on proliferation nor does it appear to involve demonstrable soluble mediators. The nature of failure of MLC between TH-2 subpopulations appears to be dependent on the exppression of some product of the K, I regions of the H-2 locus. Possible mechanisms by which a B-cell-like helper cell triggers TH-2 proliferation are discussed terms of the present knowledge of specific alloantigen receptor on T and B cells, and the immunoglobulin Fc region receptors on T cells.

Mitogen-initiated synthesis and secretion of T cell growth factor(s) by a T-lymphoma cell line.

Of 12 T-lymphoma cell lines investigated, one line, EL-4 azgr (Thy-1+, Lyt-1+, Lyt-2-3-,Ia-, T-200+, sIg-, and FcR-) and, to a lesser extent, the parental cell line, EL-4, produced T cell growth factor(s) (TCGF) when stimulated by the T-cell mitogen concanavalin A (Con A). Induced production of TCGF-E was detected by 6 h and maximal at 18-24 h. Purified TCGF-E from this source had an approximately 30,000 mol wt and the biological activity of TCGF produced by whole spleen cells, including: augmentation of T cell-mitogen responses, cytotoxic T lymphocyte (CTL) proliferation support dependence, augmented generation of CTL, lack of strain specificity, and failure to stimulate resting T cells. TCGF-E is neither synthesized or secreted by this lymphoma cell line unless stimulated by Con A. X-irradiation up to 7,000 rad failed to inhibit synthesis and secretion. These observations have a practical application in providing a relatively homogeneous clonal cell product for T cell culture support and for structural and functional studies of the TCGF molecule(s). They suggest also a model for examining mechanisms of triggering production and secretion of a regulatory molecule that controls T cell functions.

Cellular recognition by mouse lymphocytes in vitro. I. Definition of a new technique and results of stimulation by phytohemagglutinin and specific antigens.

The media and culture conditions required for in vitro stimulation of mouse lymphoid cells are described. The medium was arginine-rich and contained heat-inactivated human serum. A component of the human sera necessary for stimulation of the cells was a natural mouse cell agglutinin, which affected both background stimulation and the degree of induced stimulation with phytohemagglutinin (PHA). Absorption of the agglutinin from the human serum rendered the medium incapable of sustaining DNA synthesis in the presence of PHA. The response to PHA of mouse spleen and thymus cells was age-dependent and, although this response was not present at birth, it rapidly rose to adult levels. Spleen cells from mice immunized with bacillus Calmette-Guérin (BCG) or sheep erythrocytes (SRBC) showed increased in vitro reactivity to added purified protein derivative (PPD) or SRBC stroma, dependent on the time of immunization. The dose response curve for the SRBC stroma stimulated, immune spleen cells is compatible with a theory of cell to cell interaction being necessary for an in vitro reaction to antigen. The possible role of the mouse cell agglutinin (AMLG) is discussed.

Quantitative studies of the immunoglobulin sequence in the response of the rabbit to a somatic antigen.

Employing a sensitive and immunoglobulin-specific assay method based upon antiglobulin augmentation, quantitative and qualitative aspects of the primary and secondary response of the rabbit to Salmonella typhimurium O antigens have been evaluated. These studies examine the validity of the method of assay for detecting and measuring gammaG- and gammaM-antibodies produced in response to whole organisms or its lipopolysaccharide. The results show that during the primary response gammaG-antibodies, not detectable by usual techniques, are produced in a pattern similar to that reported in animals stimulated by other classes of antigens. Moreover, the gammaG response following reinjection is characteristic of a secondary-type response. In contrast, gammaM-antibody levels after both primary and secondary stimulation rose equally to levels between 1 and 4 mg/ml. Despite increased sensitivity of detection and quantitative estimates of the actual molar concentration of each immunoglobulin, the minimal interval between gammaM and gammaG appearance in serum was not less than 1.5 days. The variable degree of augmentation of agglutination by antiglobulin reagent found during the immune response severely limits the quantitative usefulness of the methods developed. However, the data suggest that qualitative changes in anti-O antibodies interpretable as changes in avidity occur regularly during the immune response.

Avena magna: An Important New Tetraploid Species of Oats.

Avena magna is a new tetraploid species morphologically similar to the hexaploid A. sterilis, having a high concentration of protein, large caryopses, and outstanding resistance to crown rust. One genome in A. magna appears homologous to the As genome present in hexaploid, tetraploid, and one group of diploid species. Avena magna is a possible ancestor of cultivated oats.

The association of geographic atrophy and decreased renal function in patients with age-related macular degeneration.

PurposeThe purpose of the study was to investigate the association between area and presence of geographic atrophy (GA) and renal function, as measured by glomerular filtration rate (GFR).Patients and methodsWe retrospectively identified patients aged 50-90 years who were assigned an ICD-9 diagnosis code for age-related macular generation (AMD) between January 2012 and January 2016. Patients met inclusion criteria if they had at least one macular spectral domain optical coherence tomography volume scan, one provider note, and one GFR value in the electronic medical record. Images were evaluated for the presence of GA, area of GA, drusen, and subretinal drusenoid deposits (SDD) and for subfoveal choroidal thickness (CTh) by standard criteria. Imaging findings were correlated with the most recent GFR from the patient's chart.ResultsWe identified 107 patients who met our inclusion criteria (mean age=74 years, range 50-90 years). Overall, we found a significant correlation between the presence of GA and reduced GFR (P=0.002), which was maintained even after accounting for age and other confounders. No association between GFR and GA area was found. CTh was significantly lower in patients with GA (P=0.038) and those with decreased GFR (P=0.004). Within the SDD-positive population, GA was associated with reduced GFR (P=0.007) but only trended toward significance after controlling for age.ConclusionOur study findings demonstrate an association between impaired renal function and the presence, but not area, of GA within an AMD population. These findings may shed light on common pathogenic mechanisms for these two diseases.

Mexiletine: an effective antiarrhythmic drug for treatment of ventricular arrhythmias in congenital heart disease.

The use of antiarrhythmic drugs to suppress ventricular arrhythmias in pediatric patients with a structurally or hemodynamically abnormal heart appears to improve long-term prognosis. The previously successful use of phenytoin to treat serious ventricular arrhythmias led to the investigation for an alternative antiarrhythmic agent, in the same antiarrhythmic drug class, for those patients who develop side effects or become intolerant to phenytoin's antiarrhythmic effect. Forty-two children and young adults (age range 5 months to 34 years, mean 15.5 years) were treated with mexiletine. Arrhythmias treated were ventricular tachycardia (25), ventricular couplets (8), multiform ventricular premature beats (4) and frequent uniform ventricular premature beats (5). Anatomic diagnoses included congenital heart disease (postoperative in 26, unoperated in 2), cardiomyopathy (7), no heart disease (4) and other (3). Thirty-three patients had been previously treated with 1 to 5 (mean 1.6) antiarrhythmic drugs. In the short term, ventricular arrhythmias were effectively suppressed in 30 (71%) of all 42 patients treated. During follow-up (ranging to 42 months, median 10.6), 18 (60%) of the 30 acute responders continued to have excellent control. Early suppression of ventricular arrhythmias was more effective in patients with congenital heart disease (89%) than in those with cardiomyopathy (29%) or no heart disease (43%) (p less than 0.01). Initial complexity of ventricular ectopic activity had no effect on treatment results. Of 25 patients previously treated with phenytoin, in whom alternative antiarrhythmic therapy was required, 40% had long-term arrhythmia control when treated with mexiletine. Mexiletine therapy was terminated for side effects in only five patients (12%). Mexiletine is recommended for young patients with congenital heart disease and serious ventricular arrhythmias.

Transcatheter ablative techniques for treatment of the permanent form of junctional reciprocating tachycardia in young patients.

Five patients with medically refractory incessant supraventricular tachycardia due to a posterior septal, slowly conducting accessory connection underwent transcatheter closed chest ablative treatment. The tachycardia characteristics were consistent with the permanent form of junctional reciprocating tachycardia. In each patient the ablative attempts resulted in independent interruption of either the anterograde limb (atrioventricular node-His bundle conduction) or the retrograde limb (accessory connection) of the tachycardia circuit. Permanent retrograde pathway ablation was achieved in only one patient and followed separate permanent transcatheter His bundle ablation. In three of the other four patients the ablation attempt caused temporary interruption of retrograde conduction. Each patient had improved control of tachycardia related to the ablation attempt. Of the five patients, four required pacemaker implantation. With further refinements, selective ablation of the retrograde limb of the tachycardia circuit may be possible. This experience confirms the anatomic independence of the anterograde and retrograde limbs of the tachycardia circuit.

Flecainide acetate for resistant arrhythmias in the young: efficacy and pharmacokinetics.

Drug efficacy and pharmacokinetics were assessed in 63 patients, aged 5 days to 30 years (mean 8 years), who received flecainide acetate for control of resistant arrhythmias. Doses of flecainide ranged from 59 to 225 mg/m2 body surface area per day (mean 141) in divided doses every 8 to 12 h and serum trough levels ranged from 0.10 to 0.99 micrograms/ml (mean 0.36). Flecainide controlled or partially controlled arrhythmia in 53 (84%) of the 63 patients: 7 of 7 patients who had the permanent form of junctional reciprocating tachycardia, 12 of 13 who had an atrial ectopic tachycardia, 10 of 10 who had ventricular tachycardia and 18 of 25 patients who had reentrant supraventricular tachycardia. Five of seven patients who had the latter arrhythmia were unsuccessfully treated with flecainide. They had Wolff-Parkinson-White syndrome and developed asymptomatic, incessant, slower orthodromic reciprocating tachycardia while receiving the drug. Transient blurred vision was reported in three patients and two patients had transient hyperactivity. No significant hemodynamic side effects were seen in any patient. Twenty-five patients underwent oral pharmacokinetic investigation. Young infants (less than 1 year of age) had a mean plasma elimination half-life (t 1/2) approximating that (11 to 12 h) found in older children and healthy adults; children aged 1 to 12 years had a shorter mean t 1/2 of 8 h. Dosing schedules based on milligrams per square meter body surface area correlated better with plasma flecainide levels than did dosing based on milligrams per kilogram body weight.(ABSTRACT TRUNCATED AT 250 WORDS)

Ventricular arrhythmias and sudden death in children.

In children, sudden death related to ventricular arrhythmias occurs virtually always in a patient with an abnormal heart. Therefore, children with ventricular tachycardia should be thoroughly investigated by anatomic cardiac catheterization and possibly electrophysiologic study. Sudden death may occur in a patient who had been relatively asymptomatic. This especially occurs in patients after repair of congenital heart disease. The patient may also never have had documented ventricular tachycardia, although most have had at least premature ventricular complexes on a Holter monitor recording. Finally, sudden death related to ventricular arrhythmias can often be prevented with vigorous medical and surgical therapy.

Fetal ventricular pacing for hydrops secondary to complete atrioventricular block.

The advent of ultrasound recording has expanded the capabilities for treatment of the fetus in utero. The diagnosis of specific disease processes has allowed for prenatal intervention by new techniques designed to improve fetal survival. The application of ventricular pacing in a hydropic fetus with complete atrioventricular (AV) block is reported. Complete AV block resulted from maternal collagen vascular disease. The application of ventricular pacing was to allow for further in utero development and for reversal of hydrops fetalis after improvement in cardiac output. Despite fetal death 4 hours after placement of the ventricular pacing lead, this procedure when applied earlier in the development of hydrops may allow for fetal survival. Ventricular pacing was accomplished without apparent trauma to mother or fetus and no evidence of fetal injury was seen at necropsy. Therefore, in the fetus who would otherwise die in utero before the point of viability ex utero, fetal ventricular pacing may be a rational alternative to current observation.

Incessant ventricular tachycardia in infants: myocardial hamartomas and surgical cure.

Infants with incessant ventricular tachycardia (occurring greater than 10% of the day) have generally been described in pathologic studies. This report describes 21 patients with incessant ventricular tachycardia present greater than 90% of the day and night; the age at diagnosis ranged from birth to 30 months (mean 10.5 months). The most common clinical presentation was cardiac arrest (11 patients, in 5 after digitalis for presumed supraventricular tachycardia); another 6 patients had congestive heart failure and 4 were asymptomatic. Three patients had coexisting Wolff-Parkinson-White syndrome. The rate of incessant ventricular tachycardia ranged from 167 to 440 (mean 260 beats/min) and the QRS duration from 0.06 to 0.11 second. The most common electrocardiographic (ECG) pattern (10 of 21) was right bundle branch block with left axis deviation, but other right and left bundle branch block patterns were observed. Conventional and investigational antiarrhythmic agents (nine patients received amiodarone) failed to eliminate incessant ventricular tachycardia in all. Electrophysiologic studies localized incessant ventricular tachycardia to the left ventricle in 17 (to the apex in 2, the free wall in 9 and the septum in 6) and to the right ventricular septum in 4. No structural abnormalities were found on the echocardiogram or angiocardiogram. All 21 patients had surgery at an age of 3.5 to 31 months (mean 16). In 15 a tumor was found: 13 myocardial hamartomas (9 discrete, 4 diffuse throughout both ventricles) and 2 rhabdomyomas (1 multiple). Myocarditis was found in one patient (the oldest). In four, only myocardial fibrosis was found; results of one biopsy were normal.(ABSTRACT TRUNCATED AT 250 WORDS)

Stress testing for comparison of synthetic patch aortoplasty with resection and end to end anastomosis for repair of coarctation in childhood.

Excellent clinical results have been achieved by both resection with end to end anastomosis and synthetic patch aortoplasty for the repair of coarctation of the aorta in older children. Increasing experience with exercise stress testing in the postoperative evaluation of patients with coarctation has allowed the discovery of less obvious differences between the two procedures. To evaluate these differences further, the stress tests of 50 postoperative patients who underwent coarctation repair were reviewed: 26 patients with end to end anastomosis and 24 with synthetic patch aortoplasty. Twenty normal control subjects were similarly exercised. Systolic blood pressure in the arm and leg was evaluated before and after the test. Heart rate, electrocardiogram and arm blood pressure were monitored during the test. The mean arm systolic blood pressure was higher at all points of measurement in the patients who underwent repair by end to end anastomosis than in the group who underwent patch aortoplasty. These systolic pressure differences reached statistical significance only for standing arm blood pressure before exercise (p less than 0.05) and for supine arm systolic blood pressure immediately after exercise (p less than 0.01). There was no difference in arm-leg pressure gradient between the two study groups before exercise; however, after exercise the group with end to end anastomosis had significantly higher arm-leg pressure gradients (p less than 0.001). Significant differences between the two types of repair not apparent at rest were found immediately after exercise. The long-term prognostic importance of an exercise-induced arm-leg blood pressure gradient remains to be determined. However, exercise stress testing is sensitive in demonstrating these differences.

Prevention of sudden death after repair of tetralogy of Fallot: treatment of ventricular arrhythmias.

The majority of sudden deaths after repair of tetralogy of Fallot have been presumed to be due to ventricular arrhythmia; however, it remains to be demonstrated that antiarrhythmic medication reduces the incidence of sudden death. Since 1978, ventricular arrhythmias have been treated aggressively; these include any ventricular arrhythmia on routine electrocardiogram and more than 10 uniform premature ventricular complexes per hour on 24 hour electrocardiogram. A review was undertaken of 488 patients followed up for more than 1 month after repair of tetralogy of Fallot (mean follow-up time 6.1 years); 13.5% had ventricular arrhythmia on routine electrocardiogram. Ventricular arrhythmia appeared from 2 months to 21 years postoperatively (mean 7.3 years). Ventricular arrhythmias were significantly (p less than 0.01) related to: longer follow-up duration, older age at follow-up, older age at operation and higher postoperative right ventricular systolic and end-diastolic pressures. Ventricular arrhythmia on routine electrocardiogram occurred in 100% of those who later died suddenly compared with 12% of those who did not die (p less than 0.01). Treatment for ventricular arrhythmia was given to 46 patients and considered "successful" if there were fewer than 10 uniform premature ventricular complexes per hour on 24 hour electrocardiogram. A successful drug was found in 44 of the 46: 30 of 34 given phenytoin, 6 of 9 given propranolol, 1 of 7 given quinidine, 1 of 2 given disopyramide, 8 of 9 given mexiletine and 4 of 5 given amiodarone.(ABSTRACT TRUNCATED AT 250 WORDS)