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Background and Objectives Transfusion-associated graft-versus-host disease (TA-GVHD) is a serious, potential complication of blood transfusion that is essentially prevented by blood product irradiation. Blood product irradiation is currently performed using gamma irradiation. X-ray irradiation is an alternative that has certain advantages: an X-ray irradiator is less expensive and does not have a radioactive source. However, the biochemical effects of X-ray irradiation on red blood cells (RBCs) have not been well characterized. The primary purpose of our study was to compare the effects of X-ray irradiation with gamma irradiation on RBC membrane permeability. A secondary purpose was to verify that X-ray irradiation has the same effect on lymphocytes as gamma irradiation. Materials and Methods Ten RBC units were split into two portions; five units of each were then irradiated with either gamma or X-ray at a dose of 35 Gy or 25 Gy, respectively. Free plasma haemoglobin and extracellular potassium levels were measured in each unit over time. Another unit was divided into three parts for lymphocyte studies; one part was not irradiated as a control and the other two received 25 Gy of irradiation via X-ray or gamma irradiation, respectively. Results Minimally increased free plasma haemoglobin was found only in X-ray-irradiated units after 25 Gy compared to gamma-irradiated units. However, extracellular potassium levels were not significantly different in these groups. Extracellular potassium levels were slightly higher only in gamma irradiated units at 35 Gy. X-ray and gamma irradiation were equivalent in affecting lymphocyte function. Conclusions Small differences in RBC membrane permeability are found between gamma-irradiated and X-ray-irradiated units. However, these differences are not likely to be clinically important.

A neural network for calculating the correlation of a signal with a Gaussian function is described. The network behaves as a Gaussian filter and has two outputs: the first approximates the noisy signal and the second represents the filtered signal. The filtered output provides improvement by a factor of ten in the signal-to-noise ratio. A higher order Gaussian filter was synthesized by combining several Hermite functions together.

A method for obtaining the correlation of two Hermite neural networks is developed. The method is based on the fact that a Hermite function is unchanged by the Fourier transform, which allows an expression for the correlation to be obtained directly from the network weights without the need for the Fourier transform. Comparative results with other neural network correlation methods are presented on simulated radar signals.

Vaccination has been proposed for the prevention of disease due to enterohemorrhagic Escherichia coli (EHEC), but the immune response following human infection, including the choice of potential antigens, has not been well characterized. To study this, sera were obtained from five pediatric patients with acute diarrhea caused by E. coli O157:H7 0, 8, and 60 days after hospitalization. These sera were used to examine the immune response to four different EHEC virulence factors: Tir (translocated intimin receptor, which is inserted into the host cell membrane), intimin (bacterial outer membrane protein which binds to Tir), EspA (secreted protein which forms filamentous structures on EHEC surface), and EspB (inserted into the host membrane and cytoplasm). The response to O157:H7 lipopolysaccharide was also examined. Sera were assayed against purified recombinant proteins using immunoblot analysis and by enzyme-linked immunosorbent assay to determine the sera's titers to each of the antigens in all patients. We found that there was little reaction to EspA, EspB, and intimin in the acute-phase sera, although there was some reactivity to Tir. By day 8, titers of antibody to all four virulence factors were present in all patients, with a very strong response against Tir (up to a titer of 1:256,000), especially in hemolytic-uremic syndrome patients, and lesser strong responses to the other three antigens. The titer to the antigens 60 days after hospitalization was decreased but was still highest for Tir. These results suggest that there is a strong immune response to Tir, and to a lesser extent to the other three virulence factors, following EHEC disease, indicating that these bacterial molecules are potential vaccine candidates for preventing EHEC disease. They also suggest that bacterial virulence factors that are inserted into host cells during infection by type III secretion systems (Tir or EspB) are still recognized by the host immune response.

A two-dimensional solid state LDA system was developed for the measurement of turbulent flow in a test journal bearing. Visible laser diode and avalanche photodiode were employed in the design of the LDA optics and optical frequency shifts were used to separate signals and remove directional ambiguity. Special techniques were tested to minimize the influence of the diffused light from the shaft surface and make valid measurements near the walls. Two-component measurements were made in a journal bearing with a minimum gap of 0.49 mm and velocity profiles under different Reynolds numbers were obtained.

Recently a model of Reynolds stress in turbulent lubrication theory was proposed by Tieu and Kosasih (1992) based on a modified Van Driest mixing length formula. It was developed from a study of Poiseuille flow velocities in narrow gaps. As a continuation of that study, this paper describes an investigation into fluid velocities in fluid film bearings. Experimental velocity profiles of planar flows in various film geometries are compared with the profiles calculated using the mixing length model in the transition-turbulent regime. Excellent agreements have been attained, confirming the validity of the formula in the superlaminar theory. The effects of Reynolds number and pressure gradient on nonplanar velocity profiles are also presented.

PURPOSE Curative therapy for multiple myeloma continues to be an elusive goal. This report discusses the Northern California Oncology Group (NCOG) phase I and II trial in high-tumor-burden disease that used a strategy that consisted of induction chemotherapy (vincristine, melphalan, cyclophosphamide, and prednisone [VMCP]) for eight cycles followed by sequential hemibody radiation therapy (RT) and subsequent chemotherapy for an additional eight cycles. PATIENTS AND METHODS Seventy-two previously untreated stage III myeloma patients were entered onto the study. Sixty-nine received induction chemotherapy, 40 received induction chemotherapy and hemibody RT, and 23 received induction chemotherapy, hemibody RT, and consolidative chemotherapy. RESULTS Twenty-two complete responses (CRs) were obtained by induction chemotherapy, with four additional CRs after RT and consolidative chemotherapy. Nineteen patients developed grade 4 hematologic toxicity primarily after upper hemibody RT. Eight of these developed long-standing neutropenia or thrombocytopenia. Median survival of the group was 134 weeks, which was not significantly different from other approaches. CONCLUSIONS Hemibody RT can be combined with chemotherapy as induction therapy and can be safely administered in a community setting. However, as administered here no survival advantage was demonstrated.

A visible wavelength solid-state LDA is described. The instrument has been designed for measurements in narrow gaps. Poiseuille and Couette flow measurements are demonstrated.

Kernel regression is one model that has been applied to explain or design radial-basis neural networks. Practical application of the kernel regression method has shown that bias errors caused by the boundaries of the data can seriously effect the accuracy of this type of regression. This paper investigates the correction of boundary error by substituting an asymmetric kernel function for the symmetric kernel function at data points close to the boundary. The asymmetric kernel function allows a much closer approach to the boundary to be achieved without adversely effecting the noise-filtering properties of the kernel regression.

Background and Objectives Platelet function abnormalities have been reported in blood donors who have not consumed aspirin. Our objective was to identify factors other than aspirin that may contribute to impaired platelet function in qualified volunteer blood donors. Materials and Methods Blood samples were obtained from 24 donors following routine blood donation. Donors completed a study questionnaire that included questions about recent food consumption, medication and medical history. Platelet activation was measured using monoclonal antibodies and flow cytometry. CD62P expression and PAC-1 binding on platelets were used as indicators of platelet activation. Platelet function was measured on a platelet function analyser (PFA-100®) using both collagen/epinephrine (cEPI) and collagen/ADP (cADP) cartridges. Results Fifty-four per cent of donors (13 of 24) had normal platelet function. Thirty-eight per cent (nine of 24) had prolonged cEPI closure times, of whom four (17%) had no cEPI closure (> 300 seconds). No closure was associated with aspirin use (two donors) or chocolate consumption (two donors) before donation. Two donors (8%) had either a shortened cEPI or cADP closure time. Conclusions Platelet dysfunction in qualified blood donors is underestimated. Platelet function screening can identify donors with diet-related platelet dysfunction or with poor recollection of aspirin use.

A low-expression-class methicillin-resistant Staphylococcus aureus strain (strain 9302-2) was sent to 207 laboratories as part of the bacteriology component of the Laboratory Proficiency Testing Program of Ontario. An incorrect (susceptible) result was reported by 16 of 76 (21%) of laboratories that used the National Committee for Clinical Laboratory standards disk test, whereas 1 of 104 laboratories that used other methods reported an incorrect result (P < 0.05). Experiments showed discrepancies in the disk test results given by Mueller-Hinton agars from three manufacturers. We advise that laboratories should use a low expression-class methicillin-resistant S. aureus isolate as a control for the National Committee for Clinical Laboratory Standards disk test.

Flow cytometry assays, which measure CD69 activation and intracellular cytokine production, have been used to measure peripheral blood lymphocyte (PBL) responses to in vitro antigen exposure. In the present study, we show that, in healthy individuals and immunosuppressed kidney transplant recipients, CD69 expression and intracellular cytokine production by peripheral blood T cells compare favorably to thymidine uptake as a measure of PBL response to alloantigen in mixed leukocyte culture (MLC). Heparinized whole blood from 23 healthy individuals was incubated for 24-48 h with 3rd party allogeneic monocytes; blood from twelve kidney transplant recipients was incubated with monocytes from their kidney donor and with monocytes from unrelated individuals. The percentage of T cells expressing surface CD69 or intracellular IL-2 or IL-4 was determined by 3-color flow cytometry. We identified 5 donor-specific response patterns in our kidney transplant group. One transplant recipient was hyporesponsive; his cells did not express CD69 or produce IL-2 in response to either donor or 3rd party allogeneic cells. All other transplant recipients expressed CD69 and IL-2 in response to 3rd party allogeneic cells. Two had no response to donor cells (donor-specific hyporesponsiveness), three had donor-specific anergy (CD69 expression without cytokine production in response to donor cells), five had a donor-specific Thl response (CD69 expression and IL-2 production in response to donor cells), and one had a donor-specific Th2 response (CD69 expression and IL-4 but not IL-2 production in response to donor cells). Rapid measures of donor-specific hyporesponsiveness such as CD69 activation antigen expression and intracellular cytokine production may prove valuable in monitoring lymphocyte function and aid in the long-term management of kidney transplant recipients.

Our group has a long standing interest in the immune abnormalities present in patients with multiple myeloma. In particular the poor primary immune response to new antigen challenges exemplified by recurrent upper respiratory infections. This aberrant physiology has an in vitro correlate of diminished levels of serum immunoglobulins other than the monoclonal protein which marks the expression of the plasma cell tumor. Attempts to define the mechanisms of this immunodeficiency traditionally use the manipulation of production of immunoglobulins by peripheral blood mononuclear cells stimulated by pokeweed mitogen (PWM). In this system the inhibition of Ig production by added cell populations or culture supernatants is determined. Recently the use of Staphylococcal Aureus protein (SAC) as a stimulant proved a satisfactory system as well.

Diverse indicators were used to examine medical research in Chile. According to participation in Fondecyt projects, it was concluded that 420 persons were involved in medical sciences research and that 41.9% of them had no postgraduate title. During the eighties, there was a clear aging of the community dedicated to medical research. The scientific productivity was assessed through the inventory of publications in indexed journals. During the eighties, the number of articles in Clinical Medicine and Biomedical Sciences increased two-fold. The University of Chile published the greatest number of articles, followed by the Catholic University and, with a remarkable lower amount, other Universities. The impact of the published articles in Clinical Medicine and Biomedical Sciences was calculated from the citations that they generated from 1986 to 1988. Likewise, the articles registered in 1985 with the higher citation indexes were identified.

We report the results of a province-wide quality control program in which five methicillin-resistant Staphylococcus aureus strains were circulated to all Ontario laboratories (hospital, private, and public health laboratories) on nine occasions between 1980 and 1989. The level of expression of methicillin resistance in each of the isolates was determined by performing viable colony counts on serial dilutions of methicillin in agar, and each isolate was assigned to an expression class according to previous published criteria (A. Tomasz, S. Nachman, and H. Leaf, Antimicrob. Agents Chemother. 35:124-129, 1991). Over this time there was an improvement in the performance of laboratories in the recognition of three strains that were relatively easy to detect (strains B, C, and E). These strains were of expression class II, and 98% of laboratories reported correct identifications in 1986. Performance in identifying two strains (strains A and D) of expression class I remained poor. Strain A was circulated in two surveys in 1987 and 1989, and laboratories were sent a questionnaire requesting details of the methods used in those two surveys. The methods used by the laboratories were classified into three categories: disk diffusion, single-plate screening by agar incorporation, and automated methods, which included premanufactured MIC panels. Between the 1987 and 1989 surveys, there was no change in the performance of the disk diffusion test (60% correct on both occasions), but there was improvement in the sensitivity of the agar incorporation test (36% correct in 1987 and 84% correct in 1989) and in automated methods (43% correct in 1987 and 79% correct in 1989). Over a decade, there was overall improvement in the performance of laboratories in detecting easy-to-detect strains, but there were difficulties in detecting organisms of low expression class, and an organism of very low expression class should be designated as a control organism for routine testing of methicillin-resistant s. aureus isolates.

A continuously growing plasma cell line has been established from the bone marrow of a multiple myeloma patient. Initial growth of the cells was dependent on the presence of bone marrow stromal cells. Following initial outgrowth the cells were maintained by transfer onto non-autocthonous bone marrow stromal cultures. Following approximately one year of continuous growth, a subline was derived which could be grown independently of feeder cells. These stromal-cell-independent myeloma cells nevertheless retained dependence for a growth factor present in stromal-cell-conditioned media. The relevant factor in the conditioned media was determined to be interleukin-6 (IL-6). The cells also ultimately became independent of the conditioned media. These latter cells were shown to contain mRNA for IL-6 and eventually began to secrete IL-6. This cell line has thus progressed from complete dependence on stromal cells to IL-6-dependent growth in the absence of stromal cells to complete self sufficient growth. This in vitro progression may reflect an in vivo pattern of myeloma development.

A continuously growing, in vitro cell line with plasma cell characteristics has been established from the bone marrow of a patient with multiple myeloma. Surface marker characterization of the cells revealed a combination of markers normally associated with different developmental stages in the B cell lineage. The cell secretes immunoglobulin at a relatively low rate. The cell also expresses CD5 and secretes a factor which suppresses in vitro mitogen and antigen induced immunoglobulin synthesis by normal PBL.

The application of a solid-state laser doppler anemometer (LDA) to laminar flow occurring in a 175 μm channel is reported. Details of the optical system are described.

A low-noise receiver for a miniature visible laser diode anemometer is reported. Application of the anemometer to measure laminar oil flow in a narrow channel is described.

Patients with myeloma have a depressed capacity to respond to antigenic challenge. Studies in this laboratory have previously described an unclassified lymphoid cell which binds human erythrocytes coated with human immunoglobulin G (IgG) anti-D antibody (EA) as important in the inhibition of Ig synthesis in myeloma patients. Using monoclonal antibodies, two-color fluorescence studies, and flow cytometry, we characterized this EA cell as a Leu-1+ (cluster designation (CD) 5), Leu-12+ (CD 19), Leu-16+ (CD 20), B2+ (CD 21), Leu-14+ (CD 22), and HLA-DR+ B cell. The cell was negative for antibodies to Leu-2 (CD 8), Leu-3 (CD 4), Leu-4 (CD 3), Leu-5 (CD 2), Leu-7, Leu-8, Leu-11 (CD 16), Leu-M1 (CD 15), Leu-M3, and CALLA (CD 10). This profile is consistent with a Leu-1+ B cell and excludes a T cell, natural killer cell, and monocyte. Comparison of the relative role of these cells to the role of monocytes in the suppression of pokeweed mitogen-stimulated Ig synthesis was determined in serial studies on 19 myeloma patients. The mean (+/- SEM) percentage of inhibition of Ig synthesis by monocytes from stage I myeloma patients was 14 +/- 2.2%, from stage II patients was 37 +/- 3.5%, and from stage III patients was 51 +/- 4.7%. Inhibition of Ig synthesis by Leu-1+ EA cells was 46 +/- 1.5%, 48 +/- 1.6%, and 43 +/- 3.7% in stage I, II, and III patients, respectively. Immunosuppressive B cells are an important component of inhibition of Ig synthesis in the immunodeficiency of myeloma.

Antisera made to prototype serogroup B strains of Neisseria meningitidis were used to serotype, by agar gel double diffusion, 262 meningococcal serogroups B and C strains isolated in Canada. The strains included 93 from patients and 169 from carriers. Serotype 2 was associated with 39 of 75 (52%) of group B strains and 14 of 18 (77.8%) of group C strains isolated from patients. The group B strains were mainly (87.2%) serotype 2b, while the majority (92.2%) of group C strains was serotype 2a. Other serotypes (including a new provisional serotype) represented 25.3 and 5.5% of groups B and C strains, respectively. The new serotype accounted for 13% of the group B strains. Approximately 23% of the strains isolated from patients were nontypable. The distribution of serotype 2, nontype 2 (other serotypes), and nontypable strains isolated from carriers was 2.1, 36.6, and 61.3%, respectively, for group B meningococci and 22.2, 29.6, and 48.2%, respectively, for group C meningococci. Serotype 11 was the most prominent of the strains isolated from carriers. Approximately 7% of all the strains were multiple serotypes. Serotype 2 is an important virulence marker associated with meningococcal groups B and C disease in Canada, with serotypes 2a and 2b being markedly associated with groups C and B meningococcal disease, respectively.

A patient is described who suffered from severe chronic asthma complicated by repeated chest infections. She developed a large gelatinous pseudotumour of the lung which was found to be caused by Bacillus sphaericus.

The binding of Staph A protein to murine immunoglobulins has previously been thought to be restricted to the IgG2 and IgG3 classes. In this study five IgM proteins were assessed for binding, and of these one showed marked Staph A binding. Pepsin digestion of this IgM molecule produced several different sized fragments, and the binding studies with these fragments indicated that the binding site is in the CH2 domain.

Enrichment culture with a semisolid medium increased by 6% the isolation rate of Campylobacter fetus subsp. jejuni. The semisolid enrichment medium was also used successfully as a transport medium for Campylobacter isolates. A blood agar plate streaked with Pseudomonas aeruginosa, Clostridium perfringens, and a laboratory strain of Campylobacter was a good control system for the microaerophilic atmosphere. Good growth of all three organisms indicated satisfactory conditions for culturing Campylobacter.

Lymphoid cells bearing receptors for human IgG anti-Rh antibody but not otherwise identifiable as T or B cells were isolated from the peripheral blood of human multiple myeloma patients. These cells do not demonstrate surface immunoglobulin, do not secrete immunoglobulin in culture after stimulation with pokeweed mitogen, and do not possess receptors for the third component of complement. They do not form rosettes with sheep erythrocytes, and do not respond to known T cell mitogens. These cells are not phagocytic and do not possess demonstrable nonspecific esterase. Thus, they must be considered at present, members of the "third population" lymphoid cells. These cells are active in the cytotoxic response by myeloma peripheral blood lymphocytes to myeloma plasma cells accounting for approximately 60% of such activity. They are also responsible for part of the suppression of pokeweed mitogen-stimulated immunoglobulin production, and at equivalent suppressor to target cell ratios are more active than immunoregulatory monocytes. Thus, these cells play an important, not previously widely appreciated role in the pathophysiology of the human myeloma patient.

Summary Serologic differences between γM-globulin antibodies such as Rh saline agglutinins, the α- and β-isohemogglutinins and cold and heterophile agglutinins have been demonstrated by the use of rhesus monkey antisera to individual Rh saline agglutinins and to Waldenström γM-globulins. Differences in the γM-globulin antibodies within an individual human sera also exist. Rhesus monkey antisera to γM-globulins give multiple precipitin bands with normal human serum or with isolated γM-globulins when the reactions are carried out in agar gel. This further indicates the antigenic complexity of these proteins. Immunoelectrophoretic studies in agar indicate that the primary factors in human serum responsible for the precipitin reactions with the monkey anti-Rh saline agglutinins are γM-globulins. Starch gel immunoelectrophoretic experiments confirm this and further indicate that other proteins are present in Type O sera that react with monkey antibody to γM-globulins.

Campylobacter jejuni Skirrow biotype 1, Lior serotype 8 was isolated from the appendix of an 11-year-old boy who had a 6-h history of acute abdominal pain. Histological diagnosis on the appendix section was early acute appendicitis. Dilute carbol fuchsin stain and indirect fluorescent antibody test performed on the appendix section also revealed the presence of Campylobacter sp. The patient developed a significant bactericidal antibody titer of 1,024, providing substantial clinical evidence of the pathogenicity of the isolate. This case indicated that not only may abdominal pain caused by Campylobacter enteritis mimic appendicitis, but the organism may actually be recovered from the infected appendix.

Enrichment media for isolation of therm0Philic campylobacters from stools include alkaline peptone water (1), Campy-thio (2), nutrient broth (3), semi-solid motility test medium (4), and bmcella broth (5). Campy-thio has been widely accepted in the United States and is now available commercially (BBL Microbiology Systems, USA). Our experience with a semi. solid enrichment medium indicated that it had a higher isolation rate than Campy-thio. Since a parallel study of the two media has not yet been reported, we investigated the value of enrichment culture for isolation of campylobacter from contacts of patients with eampylobacter enteritis, comparing the performance of two primary selective media and three enrichment methods.

We analyzed the clinicopathologic features of 13 patients with immunologically confirmed peripheral T-cell lymphoma. The lymphomas were classified into poorly differentiated lymphocytic, mixed cell, and large cell types. Marked morphologic heterogeneity was noted within the mixed cell and large cell categories, and the various subtypes are described. Twelve of the 13 patients received multiagent chemotherapy. Only three of the nine patients with poorly differentiated or mixed cell lymphomas achieved a complete remission, and the median survival for this group was 11 months. In contrast, all three of the treated patients with large cell lymphomas achieved a complete remission, two of whom are alive without disease (14 and 29 months, respectively). Classification of peripheral T-cell lymphomas into lymphocytic, mixed cell, and large cell types, as well as further subclassification within the heterogeneous groups, is suggested so that pathologic features of prognostic significance can be identified.

Bone marrow mononuclear cell populations were studied in 35 patients without myeloma, 39 patients with multiple myeloma, and 15 patients with benign monoclonal gammopathy. Bone marrow mononuclear cell receptors, responses to mitogens or allogeneic stimuli, and suppressive effects on in vitro peripheral blood lymphocyte (PBL) function were studied. In bone marrow cell populations from patients with untreated multiple myeloma, the percent of complement receptor-bearing cells and the pokeweed mitogen- and concanavalin A-stimulated responses were significantly greater than were those in bone marrow cell populations from patients without myeloma. Sheep red blood cell receptor-bearing cells were significantly greater in marrow populations from treated multiple myeloma patients compared to those from untreated multiple myeloma patients. Sheep red blood cell receptor-bearing cells from the bone marrow of multiple myeloma patients suppressed responses of the multiple myeloma patients' PBL's to autologous mitomycin C-treated bone marrow plasma cells and to allogeneic stimuli in one-way mixed leukocyte culture. Complement receptor-bearing cells suppressed the response to pokeweed mitogen. The presence of lymphocytes in the marrow compartment that are capable of suppressing the response of myeloma patients' PBL's to plasma cell antigens may be significant in the pathogenesis of multiple myeloma.

Seventy-four patients with multiple myeloma, 17 untreated and 57 treated, were studied to characterize their peripheral blood lymphocytes. PBL were studied for E, EAC, and EA rosette-forming cells, SIg, and Fc receptor-bearing cells. The responses to HA, Con A, and PWM were assessed as well as their ability to stimulate or to respond in a MLC. Finally, the capacity of mitogen-stimulated lymphocytes to lyse Chang cells, CRBC, and PHA-stimulated lymphoblasts was examined. These results were compared with a group of normals and patients with benign monoclonal gammopathy. In untreated myeloma patients there was a normal percentage of T cells, but an abnormal distribution of B cells as judged by a decrease in SIg-bearing cells, as well as an increase in EAC rosette-forming cells. Subpopulation analysis showed a marked increase in EAC rosette-forming cells without SIg. PHA, Con A, and PWM, and response in MLC were all normal. However, the ability to stimulate in MLC was significantly depressed. Treated myeloma patients had similar findings, except that the response to PWM was significantly depressed. The capacity of PWM-stimulated cells to lyse target cells was depressed in both groups. The results indicate that, in the peripheral blood of myeloma patients, there are populations of lymphocytes characterized by the presence of the EAC receptor without SIg, which are deficient in the capacity to stimulate an MLC response and the ability to be cytotoxic when stimulated by PWM. The results form a baseline for the study of abnormal lymphoid function in human myeloma.

The comparative adherence of seven strains of Staphylococcus epidermidis to plastic surfaces was measured by an agar overlay technique in which adherent organisms were counted by their ability to form colonies under an agar overlay. The degree of adherence to plastics decreased in the order polystyrene, polyvinyl chloride, Silastic, and polytetrafluoroethylene.

The binding of murine IgG1, IgG2a, IgG2b, IgG3 protein molecules to Staphylococcal A protein is presented. Differential elution by sodium thiocyanate gradients is described. Proteins of the IgG2 class (either a or b subclass) require between 1.5 and 2.0 M for complete elution, whereas, IgG1 proteins are fully eluted with only 0.5 M NaSCN. These differential elution patterns can be utilized to distinguish between, or prepare proteins of different Ig classes. It is proposed that this quantitative rather than qualitative distinction between Ig subclass binding to Staph A, indicates the existence of multiple binding sites per molecule, with different markers of such sites being present on the heavy chain of the different Ig classes.

Patients with myeloma have a depressed capacity to respond to antigenic challenge. Studies in this laboratory have previously described an unclassified lymphoid cell which binds human erythrocytes coated with human immunoglobulin G (IgG) anti-D antibody (EA) as important in the inhibition of Ig synthesis in myeloma patients. Using monoclonal antibodies, two-color fluorescence studies, and flow cytometry, we characterized this EA cell as a Leu-1+ (cluster designation (CD) 5), Leu-12+ (CD 19), Leu-16+ (CD 20), B2+ (CD 21), Leu-14+ (CD 22), and HLA-DR+ B cell. The cell was negative for antibodies to Leu-2 (CD 8), Leu-3 (CD 4), Leu-4 (CD 3), Leu-5 (CD 2), Leu-7, Leu-8, Leu-11 (CD 16), Leu-M1 (CD 15), Leu-M3, and CALLA (CD 10). This profile is consistent with a Leu-1+ B cell and excludes a T cell, natural killer cell, and monocyte. Comparison of the relative role of these cells to the role of monocytes in the suppression of pokeweed mitogen-stimulated Ig synthesis was determined in serial studies on 19 myeloma patients. The mean (+/- SEM) percentage of inhibition of Ig synthesis by monocytes from stage I myeloma patients was 14 +/- 2.2%, from stage II patients was 37 +/- 3.5%, and from stage III patients was 51 +/- 4.7%. Inhibition of Ig synthesis by Leu-1+ EA cells was 46 +/- 1.5%, 48 +/- 1.6%, and 43 +/- 3.7% in stage I, II, and III patients, respectively. Immunosuppressive B cells are an important component of inhibition of Ig synthesis in the immunodeficiency of myeloma.

A case of T-cell lymphoma, as defined by immunologic studies of lymph node, peripheral blood, and cultured cells, is presented. Convoluted lymphocytes were noted in the original lymph node biopsy, in cerebral spinal fluid preparations and terminally, in the peripheral blood. The prominent neuologic abnormalities representing both central and peripheral nervous system involvement were atypical. Other features included skin and testicular infiltration, leukemic transformation, and refractoriness to therapy. A somewhat similar clinical picture has been reported in other patients with diffuse, poorly differentiated lymphocytic lymphomas, some of which have proven to have a T-cell origin. T-cell lymphoma may represent a distinct clinical entity that calls for modifications of our traditional therapeutic approaches.

Type D retrovirus was isolated from rhesus macaques with simian acquired immunodeficiency syndrome (SAIDS) and transmitted to healthy rhesus macaques with tissue culture medium containing the virus. The clinical, immunologic, and lymph node morphologic changes were observed in 9 rhesus macaques for 52 weeks after inoculation. A spectrum of clinical signs developed including early death, persistent SAIDS, and apparent remission. Animals that died or developed persistent SAIDS had characteristic lymphoid depletion, persistently depressed peripheral blood mononuclear cell (PBMC) mitogenic response, and decreased serum immunoglobulins. The SAIDS retrovirus (SRV) was recovered from PBMC of 8 of the animals after inoculation. Virus could not be recovered from PBMC of one animal in remission, but this animal developed serum-neutralizing antibodies to SRV after inoculation. Seven of the animals seroconverted to SRV after inoculation, all 9 were seronegative for human T-lymphotropic virus-III, and 5 animals tested were seronegative to human T-lymphotropic virus-I. These findings support the etiologic role of the type D retrovirus in SAIDS and further define the pathogenesis of this disease.

Simian acquired immunodeficiency syndrome is a fatal immunosuppressive disease caused by type D retroviruses such as simian acquired immunodeficiency syndrome retrovirus type 1 (SRV-1). The disease is characterized by generalized lymphadenopathy, opportunistic infections, and lymphoid depletion with defects in both humoral and cell-mediated immunity. To understand how SRV-1 infection relates to the immune defect, we studied in vivo-infected lymphocytes from SRV-1-positive macaques with and without clinical signs of immunosuppressive disease. B and T helper/inducer and T suppressor/cytotoxic lymphocytes were purified by panning or by flow cytometry. Neutrophils were purified by dextran sedimentation, and platelets were purified by low-speed centrifugation. In vitro infection studies were also done with HUT78, H9, K562, rhesus lung fibroblast, rhesus monkey kidney, and bat lung cells. SRV-1 in lymphocytes or culture supernatants was detected by the induction of syncytia in cocultivated Raji cells and was confirmed by immunofluorescence, electron microscopy, or reverse transcriptase assay. We found that B and T helper/inducer lymphocytes were infected in all animals tested. The number of infected T suppressor/cytotoxic cells was generally lower than that of the other cell subsets, and not all animals in this subset had SRV-1 infections. All other cells exposed in vitro to SRV-1, except bat lung cells, were able to be infected. These findings show that SRV-1 has a broad cell tropism for lymphoid and nonlymphoid cell types.

Patients with Waldenström macroglobulinemia were studied for the presence or absence of the hyperviscosity syndrome, the relative serum viscosity value, and the calculated whole blood viscosity to identify a level at which symptoms occurred. The majority of symptomatic patients had whole blood viscosity values above 8.0 centipoises. There was a direct correlation between whole blood viscosity and relative serum viscosity, r = 0.75. One patient with central nervous system abnormalities was identified as having a high whole blood viscosity but a low serum viscosity. It was concluded that the vast majority of patients with the hyperviscosity syndrome will be identified by measuring the relative serum viscosity. In patients with central nervous system findings and a low serum viscosity, the whole blood viscosity should be determined either by direct measurement or by calculation.

Microorganisms may be resistant to antibiotics by one of four basic mechanisms: 1) they may synthesize enzymes that inactivate antibiotics; 2) target sites on the microorganisms may be resistant to antibiotic binding; 3) they may become resistant to particular antibiotics by the development of metabolic pathways which bypass the site of the antibiotic’s action; 4) they may modify their inner or outer membrane to prevent the penetration of the antibiotic to the microbial target site.

Tumor-specific antigens capable of eliciting a response from autologous lymphocytes have been described in mouse plasmacytoma systems. This paper presents evidence of similar antigens in human myeloma. Myeloma plasma cells isolated from the bone marrow of 27 patients stimulated autologous and allogeneic peripheral blood lymphocytes (PBL) from myeloma patients in mixed leukocyte culture. Plasma cells isolated fromt the bone marrow of normal patients or patients with benign plasmacytosis failed to stimulate PBL in mixed leukocyte culture. Similar results were found in passive cytotoxicity assays with the use of chinken red blood cells (CRBC) coated with 3M KC1 plasma cell extracts from myeloma patients. PBL from myeloma patients caused 30 to 80 percent chromium-51 release from tanned chromium-labeled plasma cell extract-coated CRBC targets, whereas PBL from normal patients or patients with benign plasmacytosis caused only 10 to 25 percent chromium-51 release. This study indicates the presence of material resembling tumor-specific antigens on human myeloma plasma cells. Immune response to suce antigens is elicited in autologous and allogeneic myeloma patients.

Peripheral blood leukocytes (PBL) from myeloma patients were studied for their capacity to lyse plasma cells from myeloma patients, benign monoclonal gammopathy (BMG) patients, and nonneoplastic disease patients. Plasma cells were isolated from bone marrow, labeled with 51Cr, and cultured with PBL isolated from patients with myeloma, BMG, or nonneoplastic disease, as well as normal individuals. PBL from patients with multiple myeloma demonstrated responses to autologous or allogeneic myeloma plasma cells. Optimum conditions for cytotoxic response included a responder-to-stimulator ratio of 1:1 and an effector-to-target ratio of 20:1. PBL from normal individuals or patients with BMG failed to demonstrate this response. However, PBL from BMG patients, but not normal individuals, could be induced to kill myeloma plasma cells (but not nonmyeloma plasma cells) by simultaneous stimulation with allogeneic lymphocytes and myeloma plasma cells.

A stable line of IgG K producing human plasma cells was established from a myelomatous human bone marrow using conditioned media from a rapidly metabolizing lymphoblast line, RPMI 4098. Growth in RPMI 1640 (15% fetal calf serum) at 6% CO2 promoted a 62-hour doubling time with a preferred cell concentration of 1 x 10(6)/mL. Surface marker studies showed: no receptors for sheep erythrocytes, no surface immunoglobulins, variable number of cells bearing complement receptors and 83% bearing Fc receptors. Although transmission electron micrographs demonstrated a poorly developed endoplasmic reticulum, radioimmunoassay showed 23 ng IgG and 28.7 ng Kappa were produced by 1 x 10(6) cells in 72 hours. Further, the cells are lipase, esterase and Epstein-Barr nuclear antigen negative. ASG banding showed a total chromosome number that varied from 46--49. Since the number of human plasma cell lines is limited, it is felt that this line will augment the immunobiological study of human myeloma.

Lymphoid cells bearing receptors for human IgG anti-Rh antibody but not otherwise identifiable as T or B cells were isolated from the peripheral blood of human multiple myeloma patients. These cells do not demonstrate surface immunoglobulin, do not secrete immunoglobulin in culture after stimulation with pokeweed mitogen, and do not possess receptors for the third component of complement. They do not form rosettes with sheep erythrocytes, and do not respond to known T cell mitogens. These cells are not phagocytic and do not possess demonstrable nonspecific esterase. Thus, they must be considered at present, members of the "third population" lymphoid cells. These cells are active in the cytotoxic response by myeloma peripheral blood lymphocytes to myeloma plasma cells accounting for approximately 60% of such activity. They are also responsible for part of the suppression of pokeweed mitogen-stimulated immunoglobulin production, and at equivalent suppressor to target cell ratios are more active than immunoregulatory monocytes. Thus, these cells play an important, not previously widely appreciated role in the pathophysiology of the human myeloma patient.

Reference values for T and B lymphocytes were determined on lymphocytes from canine thymus, spleen, lymph node, bone marrow, and peripheral blood by use of erythrocyte (E) and erythrocyte-antibody-complement (EAC) rosette assays, plus a direct fluorescent technique for assay of surface immunoglobulins. Numbers of T lymphocytes, indicated by E rosette formation with human erythrocytes, ranged from a low of 1% in the thymus to 13% in the peripheral blood, whereas B-lymphocyte numbers ranged from 3% (thymus) to 41% (bone marrow) and from 6% (thymus) to 36% (bone marrow), as indicated by EAC rosette formation or presence of surface immunoglobulins respectively. Stimulation of peripheral blood lymphocytes with either phytohemagglutinin or concanavalin A increased the total number of E-rosetting cells two to threefold, whereas the number of EAC-rosetting cells decreased by half. Further, the percentage of cells bearing Fc receptors increased after phytohemagglutinin stimulation. These results indicate the E rosette technique can be used to identify and to monitor a population of canine T lymphocytes.

Individuals with Down's syndrome are thought to have abnormalities of immune function. Studies to quantify the number of peripheral blood T and B lymphocytes and serum immunoglobulins in 12 individuals and 12 sex and age matched control subjects were performed. Hepatitis B antigen and antihyroglobulin antibodies as markers of possible immune dysfunction were determined. The numbers of circulating T and B cells, and the level of serum immunoglobulins in children with Down's syndrome did not differ from nonretarded control children. Circulating hepatitis B antigen and antihyroglobulin antibodies were not present. These studies indicated that quantitative abnormalities of T and B cells are not present in children with Down's syndrome. The data did not exclude the existence of qualitative abnormalities.