SIg-positive Cells Research Articles

Expression profiles of TCRβ and CD8α mRNA correlate with virus-specific cell-mediated cytotoxic activity in ginbuna crucian carp

Our previous studies have demonstrated that virus-specific cell-mediated cytotoxicity of sensitized leukocytes can be induced using clonal ginbuna crucian carp and their syngeneic cell lines. In the present study, we attempt to determine if virus-specific cytotoxic cell populations of fish express CD8α and TCRβ genes. Leukocytes from ginbuna crucian carp were separated into four fractions by immunomagnetic separation and density gradient centrifugation: Fraction A, leukocytes with a density of 1.08 g/ml (primarily lymphocytes); Fraction B, sIg-negative leukocytes with density of 1.08 g/ml; Fraction C, sIg-positive cells (primarily B-lymphocytes); Fraction D, leukocytes with a density of 1.08–1.09 g/ml (primarily neutrophils). Leukocytes in all fractions from uninfected fish do not exhibit cytotoxic activity against virus-infected syngeneic cells and weakly express CD8α and TCRβ mRNAs. In contrast, leukocytes in fractions A and B from virus-infected fish exhibit a high level of cytotoxic activity and strongly express CD8α and TCRβ mRNAs. In addition, mRNA expressions of CD8α and TCRβ in effector cells are upregulated by cocultivation with virus-infected target cells but not uninfected ones. The present study suggests that fish possess virus-specific cytotoxic cells with phenotype and gene expression pattern similar to those of CTLs in mammals.

Immature B lymphocytes from adult bone marrow exhibit a selective defect in induced hyperexpression of major histocompatibility complex class II and fail to show B7.2 induction.

Mature B lymphocytes respond to antigen receptor ligation by phenotypic changes, including up-regulation of major histocompatibility complex class II molecules and expression of B7.2, which are required for initiating and sustaining a productive interaction with T helper cells. We have previously demonstrated that neonatal B cells fail to show a similar up-regulation of class II and B7.2 expression following B-cell receptor (BCR) ligation, although these responses could be induced by other stimuli. Here we demonstrate that immature B cells from adult bone marrow exhibit even more profound defects in these responses, as they fail to up-regulate class II in response to either BCR ligation or interleukin-4. Moreover, bone marrow-derived, immature B cells could not be induced to express B7.2 either by receptor cross-linking or by lipopolysaccharide. These differences in the inducible expression of class II and B7.2 appear to be intrinsic to the B cells, as they were retained in purified populations of B-lineage cells and could not be induced in mature B cells by co-culture with bone marrow cells. Furthermore, short-term culture of bone marrow permitted B-cell maturation, which was accompanied by acquisition of responsiveness to the same stimuli as mature, splenic B cells. The inability of immature B cells to show these responses provides a molecular explanation for their reported deficiency in interacting with T cells. Failure of immature B cells to inducibly express B7.2 may also be important for the establishment of self tolerance in the B-cell compartment.

黒毛和種新生子牛における末梢血リンパ球サブクラスの経時変化

Changes in levels of lymphocyte subpopulations (CD 2 and SIg positive cells) and Tcell subsets (CD 4 and 8 positive cells) in peripheral blood of Japanese Black newborn calves were monitored from birth to day 28. Japanese Black Calves within 14 days after birth had lesser amounts of B-cells compared with adults. Calves from birth to day 28 have considerable amounts of T-cell and T-cell subsets as adults. Interestingly, percentages and numbers of CD2, 4 and 8 positive cells on day 3, and SIg positive cell on day 7 tended to decrease.

Properties of Nine Continuous B‐Cell Lines Established from Enzootic Bovine Leukosis Tumors

We established 9 cell lines from 63 tumor cases of enzootic bovine leukosis and studied their properties. Cells of all lines formed small clumps and floated in culture medium, indicating growth. Four of the 9 cell lines were surface immunoglobulin (SIg)-positive, but the remaining 5 line cells were negative for SIg or, if SIg was detected, the percentage of SIg-positive cells was very low. Tests for the properties of the cells with monoclonal antibodies to lymphocytes revealed that the established line cells are B-lymphocytes. Morphological observation also revealed that they had the morphology of B-lymphoblastic cell. The results of E and EAC rosette assay were negative, but 6 of 8 cell lines were positive for EA rosetting. All the 9 cell lines reacted with MoAb C-143, which recognizes the tumor-associated antigen (TAA) of the EBL tumor cell. All 9 cell lines produced bovine leukosis virus (BLV). These results suggest that the 9 cell lines are tumor cells derived from B-lymphocytes of EBL.

LYMPHOCYTE SUBSETS AND AUTOANTIBODIES IN PATIENTS WITH AUTOIMMUNE POLYENDOCRINOPATHY - CANDIDOSIS ECTODERMAL DYSTROPHY (APECED)

The basic derangement leading to immune destruction of endocrine glands in APECED is unknown. We studied lymphocyte subsets in 42 patients yith APECED and in 21 healthy controls. Lymphocytes were treated with monoclonal antibodies detecting pan-T (OKT3), T-helper (OKT4), T-suppressor/killer (OKT8), E-rosetting (OKT11) T-lymphocytes, and surface immunoglobulin (SIG)-positive B-lymphocytes, and analyzed by fluorescence activated cell sorter (FACS IV). The patients had higher lymphocyte counts than controls (2.71+1.20 vs 2.10+0.52 × 109/l; mean+SD; p=0.006). For all T-lymphocyte subsets studied, the dispersion of counts was wider for the patients than the controls. The mean counts were significantly different only for SIG-positive B-lymphocytes (0.50+0.28 vs 0.28+0.11 × 109/l; p=0. 0001) .The OKT4-OKT8-ratio was subnormal (>1.0) in 5 and supranormal (<3.1) in 5 patients. The deviations observed did not correlate with any indicator of the severity of APECED or presence of any component of it. Presence of circulating ovarian, testicular or placental antibodies in contrast to adrenal antibodies was associated with supranormal counts of SIG-positive cells (p=0.031, p=0.02, p=0.004, p=0.35; respectively). Presence of circulating thyroid microsomal or thyroglobulin antibodies, or parietal cell or islet cell antibodies did not correlate with any lymphocyte subset pattern. No single feature was characteristic of APECED. Presumably the lymphocyte subset deviations are secondary manifestations of APECED.

Disproportionate lymphoid cell subsets in thalassaemia major: the relative contributions of transfusion and splenectomy

The relative proportions of T-cell (OKT3-positive, OKT4-positive and OKT8-positive) and B-cell (SIg-positive) populations in peripheral blood obtained from 29 chronically transfused patients with beta-thalassaemia major were compared with those of 17 healthy controls. Changes attributable to blood transfusion and/or splenectomy are described. The percentage of OKT8-positive (T-suppressor) cells found in the thalassaemic patients increased linearly (P less than 0.001) with the number of units transfused, irrespective of splenectomy. The percentage of OKT4-positive (T-helper) cells varied inversely with increasing transfusion in nonsplenectomized patients while in those who were splenectomized no significant correlation was apparent. Thus, in both groups of patients the T4/T8 ratio declined in a transfusion-related manner. The splenectomized patients experienced a marked and persistent lymphocytosis due to an increase in the number of both T- and B-cells. When the results were expressed as percentages, the greatest increase occurred in the number of B-cells, this increase being unrelated to the number of transfusions received. None of the serum parameters usually associated with iron overload or abnormal liver function correlated with the observed increases in T-suppressor and SIg-positive cells. These findings corroborate reports that transfusion of blood products may lead to decreased T4/T8 ratios. However, none of the patients studied manifested clinical signs of acquired immune deficiency syndrome (AIDS). Accordingly, studies which define transfusion related AIDS on the basis of analyses with monoclonal antibodies must be viewed with caution.

A method of purifying sheep sIg + lymphocytes as a tool for class II MHC antigen analysis

A method is described for the purification of sheep lymphocytes carrying class II MHC antigens. After incubation of purified blood lymphocytes of anti-IgM-coated petri dishes, the adherent fraction contained 95% sIg-positive cells determined by immunofluorescence. When tested with cross-reacting anti-class II (bovine and human) monoclonal antibodies, more than 95% of these cells were positive either by immunofluorescence or cytotoxicity. This technique will permit studies of the polymorphism of sheep class II antigens.

Partial characterization of bovine leukemic cell populations

Peripheral blood lymphocytes (LL) from cows with chronic lymphocytic leukemia (CLL) have been studied using a number of surface markers. Cell populations were obtained by partial enrichment and depletion using Sephadex G-200-anti-F(ab') 2 and Sephadex G-200-anti-LL-Ig immunoadsorbent columns. It was shown that cell populations having different markers, i.e., surface immunoglobulins (sIg) and leukemia-associated antigens (LAA), are characterized by similar parameters. Thus the adherent cells obtained by both fractionation methods formed 1.1–4.0% of rosettes with sheep red blood cells, while 90.6–94.3% were sIg positive cells. The cytotoxicity test (CTT) performed with anti-B and anti-LL sera indicated that a vast majority of adherent cells reacted to the sera. Thus the adherent cells represent a population with a high percentage of B-cells.

Phenotypic heterogeneity of leukemias associated with friend MuLV infection: Studies on T-cell lymphomas and null cell leukemias in euthymic and thymus-deficient mice

The development of leukemia/lymphoma in euthymic and congenitally thymus-deficient (nude) mice infected with Friend murine leukemia virus (Friend MuLV) was investigated; both groups developed fatal leukemias within 2–4 months post-infection but the gross and micropathology of lymphoid organs, coupled with cell-surface marker studies indicated the development of two distinct forms of disease. In euthymic mice one group developed lymphosarcomas manifested by thymoma, hepatosplenomegaly and lymphadenopathy whereas the second group developed splenic leukemias manifested only by hepatosplenomegaly. Analysis of surface markers on spleen cells from mice experiencing lymphosarcomas indicated that the majority of cells were positive for Thy 1.2, Moloney cell surface antigen (MCSA), and viral-coded gp70 and p30 antigens but negative for surface immunoglobulin (sIg). Euthymic mice experiencing splenic leukemias yielded spleen cells negative for Thy 1.2, sIg, and MCSA but positive for gp70 and p30. Nude mice uniformly developed splenic leukemias, spleen cells from which were Thy 1.2, MCSA, gp70 and p30 negative, although the proportion of sIg positive cells was higher than that observed in euthymic mice experiencing splenic leukemias. No correlation between the development of lymphosarcoma vs splenic leukemia and a pattern of ecotropic and/or xenotropic MuLV expression was observed. While ecotropic MuLV expression was equivalent in both euthymic and athymic mice, euthymic mice expressed approx. 10-fold higher levels of xenotropic MuLV than nude mice, however. Collectively the data suggest that infection of mice with Friend MuLV results in the development of two possible forms of disease, lymphosarcoma involving T cells vs splenic leukemia involving B and/or null cells.

Migration of bone marrow lymphocytes in mice: immunofluorescent staining using antiallotype serum.

Bone marrow cells from CSW (Igha) mice were injected intravenously into the congeneic CWB (Ighb) mice. The lymphoid tissues of the recipient CWB mice were examined for donor type surface immunoglobulin (SIg)-positive cells using fluorescein isothiocyanate-conjugated antiallotype serum Ighb anti-Igha. Donor type SIg-positive cells were rarely found in the recipients immediately after injection. However, the donor type B cells increased in the recipient's spleen from days 1-2 and reached a plateau thereafter. The present method provides a useful means for studying the migration patterns and differentiation of bone marrow lymphocytes.

The cellular content of non Hodgkin lymphomas: a comprehensive analysis using monoclonal antibodies and other surface marker techniques.

Five samples of tonsil, 10 reactive lymph nodes and 65 consecutive cases of non-Hodgkin lymphoma (NHL) were evaluated in suspension phenotyping with the monoclonal antibodies alpha Leu-I, alpha Leu-2a, alpha Leu-3a, OKT1, OKT3, OKT4, OKT6, OKT8, W6/32, 26/114, DA-2, 2DI, J5, AN51 and OKT9 together with conventional surface marking by rosetting (E, Fc gamma, Fc mu, C3b, C3d) and staining for surface and cytoplasmic immunoglobulin (SIg, CyIg) heavy and light chain classes. The results confirm the reproductability and specificity of staining with monoclonal antibodies against T cells and T cells subsets. Evidence is presented for reactivity of alpha Leu-I antibody with SIg positive and Ia positive cells in some lymphomas (centroblastic centrocytic, lymphocytic and immunoblastic), and 2 cases showed evidence of marking with OKT3 on SIg positive cells in T cell predominant immunoblastic lymphoma. Lymphoblastic lymphomas of T cell type expressed the marker OKT6. On the basis of these results criteria for the diagnosis of T cell lymphoma are suggested. The monoclonal antibody J5, reactive with C-ALL antigen, showed variable positivity, occasionally strong in B cells in cases of centroblastic and centrocytic follicular lymphoma. Proportions of cells staining with the monoclonal antibody OKT9 showed a correlation between levels of cellular expression of transferrin (trf) receptor and the histological grade of malignancy, OKT9+ cells being elevated in high grade lymphomas, and in some cases of transforming lymphoma of low grade histological class. These results are discussed and indicate the advantage of employing a wide range of defined monoclonal reagents in the phenotypic evaluation of NHL.

Monoclonal antibody analysis of blood T-cell subsets in myasthenia gravis.

Analysis of peripheral blood T -cell subsets and B-cells in patients with myasthenia gravis was performed using monoclonal antibodies and antibody against surface immunoglobulins (SIg) in an immunofluorescent technique. We found a modest but significant decrease in percentages of OKT3- and OKT8-positive cells (thought to represent total T-cells and T-suppressor/cytotoxic cells, respectively) in myasthenics as a group. The percentage of OKT3-positive cells was significantly decrease in patients with late-onset disease (greater than 35 years old), while the percentage of OKT8-positive cells was significantly reduced in those with early-onset myasthenia (greater than 35 years old). Both thymectomized and nonthymectomized patients exhibited a decreased percentage of OKT3-positive cells. No significant difference was found between the percentages of SIg-positive cells in myasthenics and controls. Our results suggest that only modest imbalances of circulating immunoregulatory lymphocytes occur in myasthenia gravis; however, it is conceivable that the small differences observed in this study may reflect pathogenetically important reductions in a functionally distinct lymphocyte subpopulation.

Immunologic markers in the differential diagnosis of non-Hodgkin's lymphomas.

To improve the classification of the phenotypes of the various types of non-Hodgkin's lymphoma (NHL), 250 cases of NHL were analyzed with immunologic and enzyme cytochemical techniques. The results confirmed previous findings. Chronic lymphocytic leukemia of B-cell type (B-CLL) is characterized by a small amount of surface immunoglobulin (SIg), a predominance of C3d receptors, a large number of mouse erythrocyte receptors, and a low T-cell content. Chronic lymphocytic leukemia of T-cell type (T-CLL) has at least two subtypes: one shows a dot-like reaction product in cells stained with acid nonspecific esterase and the other does not. In prolymphocytic leukemia, there is a constantly high percentage of SIg-positive cells and a large amount of SIg on each positive cell; C3b receptors usually preponderate over C3d receptors; and there is a large number of IgG-Fc receptors. The surface marker phenotype of hairy-cell leukemia is similar to that of prolymphocytic leukemia except that hairy cells are devoid of C3 receptors and usually show a high density of IgM-Fc receptors. T-zone lymphoma usually contains both T cells and B cells. The T cells are capable of binding sheep erythrocytes only at 4 °C and can be identified cytologically as the tumor cells. In contrast, the B cells stem from residual follicles, which are often present in T-zone lymphoma at the time of the first biopsy. Three types of lymphoplasmacytic/-cytoid lymphoma (LP immunocytoma) are distinguished on the basis of morphologic features. The marker constellation of the lymphoplasmacytic subtype resembles that of centroblastic-centrocytic lymphoma. The lymphoplasmacytoid subtype and borderline cases between this subtype and B-CLL show the same markers as does B-CLL. The third subtype of LP immunocytoma, the polymorphic subtype, differs in its marker profile from all other types of NHL. The three types of lymphoma derived from germinal center cells resemble each other in the expression of nearly equal numbers of C3b and C3d receptors and a low percentage of IgG-Fc receptors. Centrocytic lymphoma is distunguished from centroblastic-centrocytic lymphoma by a large proportion of cells bearing SIg and C3 receptors and by the absence, or small proportion, of T cells and cells rosetting with mouse erythrocytes. Centroblastic lymphoma shows a marker profile that is similar to that of centroblastic-centrocytic lymphoma. The Burkitt type of lymphoblastic lymphoma shows a unique marker profile, with a high percentage of SIg-positive cells and no other markers. Analysis of lymphoblastic lymphoma of the convoluted-cell type (including cases of acute lymphoblastic leukemia with a focal acid phosphatase reaction) revealed four phenotypes. Cases with the first phenotype show C3 receptors (usually both subtypes) and a lack of sheep erythrocyte receptors. In cases with the second phenotype, the cells express both C3 receptors and sheep erythrocyte receptors. Cases with the third phenotype lack C3 receptors but contain cells rich in receptors for sheep erythrocytes that bind at 37°C. Nearly all of the cases with these three phenotypes are devoid of acid nonspecific esterase. Cases with the fourth phenotype lack C3 receptors, exhibit sheep erythrocyte receptors that bind only at 4°C, and show a focal acid nonspecific esterase reaction. Eight of nine cases of immunoblastic lymphoma showed SIg and were thus identified as B-cell derived. The ninth case was of T-cell type, as indicated by the capacity of the tumor cells to form rosettes with sleep erythrocytes.

Surface Immunoglobulin Receptors of Rabbit Lymphoid Cells

Rabbit lymphoid cells were stained with fluorescein-labelled antiallotype antibodies. The double layer technique was found to be more sensitive than the direct staining. Rabbit B cells are stained only via their surface immunoglobulin (sIg) receptors and not via the receptors for the Fc portion of the IgG. Peritoneal exudate macrophages do not carry sIg receptors and are stained via their Fc receptors. Removal of protein aggregates from the system and use of reagents prepared from F(ab')2 immunoglobulin fragments prevent staining of the macrophages through their Fc receptors. There was a good agreement between the percent of RABELA-positive and sIg-containing spleen cells. In appendix there were approximately 20% more RABELA-positive than sIg-positive cells.

Surface Immunoglobulin Density on Human Peripheral Blood Mononuclear Cells

AbstractThe densities of surface immunoglobulin (sIg) on peripheral blood mononuclear cells (PBM) of normals and patients with chronic lymphocytic leukemia (CLL), chronic lymphosarcoma cell leukemia (LCD, and hairy cell leukemia (HCL) were analyzed using the fluorescence-activated cell sorter (FACS). PBM were labeled with fluorescein conjugates of F(ab')2 fragments of affinity chromatography-purified anti-Fab or class-specific anti-µ, anti-δ, anti-γ, or anti-α. Histograms of relative cell fluorescence, reflecting sIg density, were prepared with the FACS. Anti-Fab-labeled normal PBM demonstrated a homogeneous low-density peak that when separated by the FACS and analyzed cytochemically consisted predominantly of monocytes, whereas brighter-staining cells were predominantly lymphocytes. Anti-µ and anti-δ labeled 9.0% and 8.5% of normal PBM, respectively, the sIg+ cells being virtually all lymphocytes. Cells labeled by anti-γ exhibited low homoge neous sIg density and consisted of more than 80% monocytes. No normal or leukemic PBM were labeled by anti-α. All sIg-positive cells (<5% monocytes) from 12 of 13 patients with CLL had very low homogeneous densities of sIg and bore sIgM, whereas cells from 9 of 13 and 2 of 13 patients bore sIgD and sIgG, respectively. Similarly, PBM from 2 patients with HCL exhibited low and homogeneous densities of sIgM, sIgD, and sIgG, whereas those from a third patient bore only sIgG. By contrast, the density of sIgM and PBM derived from 3 patients with LCL was very high; sIgD and sIgG densities varied from very high to undetectable in these patients. The different homogeneous densities of sIg on peripheral blood lymphocytes from patients with CLL, HCL, and LCL suggest that these diseases represent malignant transformation of different B-lymphocyte subpopulations.

IgA-Fc Receptors on Mouse Lymphoid Cells

Abstract Receptors specific for IgA were sought on mouse lymphoid cells by using a rosetting technique in which cells are exposed to modified ox red blood cells (ORBC) coated with IgA myeloma proteins having specificity for the modified ORBC. It was found that 4 to 6% of unseparated spleen lymphoid cells (pre-incubated at 37°C overnight) formed rosettes with MOPC 315-coated TNP-ORBC, with QUPC 52-coated dextran-ORBC, and with SAPC 10-coated arabinogalactan ORBC, whereas modified ORBC alone or modified ORBC mixed with irrelevant myelomas did not form appreciable numbers of rosettes; the cells forming rosettes were not neutrophils as shown by appropriate staining techniques. Thirteen to 22% of spleen cells also formed rosettes with IgG-coated ORBC (either IgG rabbit anti-TNP-coated TNP-ORBC or rabbit IgG anti-ORBC-coated ORBC); however, by using similar techniques, no cells forming rosettes with IgM-coated ORBC could be found. The specificity of the IgA-binding sites thus demonstrated was shown in inhibition studies: IgA antigen-antibody complexes formed near equivalence or, indeed, purified IgA were found to inhibit rosette formation with IgA-coated ORBC, whereas IgG complexes or purified IgG were found to cause no such inhibition. These inhibition studies were buttressed by depletion studies in which removal of cells binding IgG-coated ORBC with rosetting procedures was shown to have little or no effect on the percentage of cells binding IgA-coated ORBC in the residual cell population. The cells capable of binding IgA-coated ORBC are at least in part T cells in that the binding of IgA-coated ORBC is a property of nylon-column passed cells containing &amp;lt;3% sIg positive cells; in addition, 2 to 3% of thymocytes bind IgA-coated ORBC. Finally, the IgA binding sites appear to be Fc-specific in that F(ab′)2 fragments of IgA do not block IgA-rosette formation. In summary, we have identified an IgA-Fc-specific binding site on mouse lymphoid cells, including mouse T cells.

In Situ Lymphoid Cells of Mouse Mammary Tumors

Abstract Lymphoid cells were isolated from mouse mammary tumors by isokinetic gradient centrifugation. ϑ-bearing and ALS-sensitive cells were always the largest subpopulations found in these tumors. In fractions 4 to 6 from the isokinetic gradient, ϑ-positive cells ranged from 8 to 46% in autochthonous tumors and 14 to 47% in passaged tumors, and ALS-positive cells ranged from 34 to 46% in autochthonous tumors and 14 to 100% in passaged tumors. SIg-positive cells were only found in occasional tumors and, where present, averaged 2% of the separated population. Fc receptor-bearing cells averaged 15% and phagocytic cells averaged 8% of the cells found in fractions 4 to 6 of the gradient. Occasional ALS-positive and one-third of the Fc receptor-bearing and phagocytic cells also separated in fractions 7 to 10 of the isokinetic gradient. Although the individual tumors were very variable in the proportions of the various lymphoid subpopulations that they contained, this variability was a stable characteristic for at least one passage generation.

Lymphocyte surface immunoglobulin in Xenopus laevis light and electron microscopic demonstration by immunoperoxidase method

To study the distribution of surface immunoglobulin (sIg * ), splenic and thymic lymphocytes of Xenopus laevis were exposed to anti— Xenopus Immunoglobulin rabbit IgG conjugated with horseradish peroxidase (aXIg-HRP). Light microscopy of lymphocytes from 6 – 8 months old toadlets revealed that sIg was detectable on 53% splenic and 91% thymic cells. “Cap” formation concomitant with internalization of sIg upon warming was evident in both splenocytes and thymocytes. The sIg positive cells in spleen were mostly small lymphocytes, although some were plasma cells, medium- and large lymphocytes. In spleen of toadlets thymectomized at Nieuwkoop and Faber stage 45 (4 days postfertilization), more than 90% of lymphocytes expressed sIg. For the spleen, lymphocytes bearing sIg seem to be thymus-independent. * Abbreviations used in this paper: aXIg, anti- Xenopus immunoglobulin rabbit IgG; aXIg-HRP, aXIg conjugated with horseradish peroxidase; HRP; horseradish peroxidase; sIg, surface immunoglobulin.

B-Lymphocytes and T-Lymphocytes in Three Types of Bovine Lymphosarcoma 2

Lymphoid cells of peripheral blood, lymph nodes, and thymus from clinically normal cattle, cattle infected with bovine leukemia virus (BLV), and cattle with lymphosarcoma were characterized for T- and B-cell surface markers. B-cells were detected by the erythrocyte-antibody-complement (EAC) rosette test and the surface immunoglobulin (sig) immunofluorescence assay. Peripheral blood from BLV-infected cattle had a higher than normal percentage of B-cells by both EAC rosette and sig immunofluorescence assays. Lymphoid cells from tumorous lymph nodes of cattle with the adult type of lymphosarcoma had a higher than normal percentage of sig-bearing cells, but in the same cell preparation the EAC rosette-positive cells were fewer than sig-positive cells. T-cells were detected by the erythrocyte rosette test. The percentage of T-cells by this test in lymph nodes of adult type lymphosarcoma was lower than that in normal cattle. A distinctly lower than normal percentage of lymphocytes could be characterized as either B- or T-cells in lymph nodes thymus, and peripheral blood from the calf type and thymic type of lymphosarcoma.