High-grade T-cell lymphoma complicating B-cell chronic lymphocytic leukemia: an unusual manifestation of "Richter's syndrome".

Approximately 3% of patients with B-cell chronic lymphocytic leukemia (CLL) develop a high-grade large-cell lymphoma consistent with Richter's Syndrome. In most cases, these lymphomas are of B-cell origin and are believed to arise by clonal evolution from the CLL cells. We present a case of a patient with a 10-year history of B-CLL who developed an aggressive large-cell lymphoma, confirmed by immunophenotype to be of T-cell origin. We suggest that in patients with CLL, immunodysregulation can result in the proliferation of T cells, which may mutate and result in the development of a new malignant clone.

Peripheral T cell lymphoma in a patient with common variable immunodeficiency disease: case report and literature review.

This report documents the occurrence of a peripheral T cell lymphoma arising in the bone marrow and liver of a patient with common variable immunodeficiency disease. The T cell origin of this lymphoma was demonstrated by immunohistochemical phenotyping and gene rearrangement studies and was not associated with EBV infection of the lymphoma cells. The frequency and characteristics of lymphomas complicating CVID are reviewed.

The effect of aging on the immune response: influence of phosphatidylcholine-containing lipid on IgD-receptor expression and antibody formation.

It was reported previously that IgD-receptors (IgD-R) are expressed on both CD4+ and CD8+ human T cells and CD4+ murine T cells after exposure to oligomeric IgD, certain cytokines, or various pharmacological agents, as shown by rosetting with IgD-coated erythrocytes. Enhancement of antibody production is observed in mice after injection of oligomeric IgD and is mediated by these IgD-R+ T cells, while injection of monomeric IgD inhibits both IgD-R upregulation and augmentation of antibody responses induced by simultaneously injected oligomeric IgD. The effects of oligomeric IgD on IgD-R upregulation are lacking in aged mice. However, the oligomeric IgD induced enhanced antibody production can be transferred to aged mice with IgD-R+ T cells from young donors suggesting that the environment of the aged mouse supports the effector function of IgD-R+ T cells. We now report, in addition, that exposure to phosphatidylcholine (PC) and a PC-containing lipid mixture, AL721, is effective in causing IgD-R upregulation on T cells from both young and aged mice, and young humans. This effect can also be demonstrated in mice in vivo after administration of AL721. Moreover, this agent causes a two-fold enhancement of antibody production, as measured by PFC/spleen, to 4-hydroxy-5-iodo-3-nitrophenyl(acetyl)-Brucella abortus (NIP-BA) and NIP-horse red blood cells (RBC) in young and aged mice. There is no difference in the baseline membrane fluidity of lymphocytes from aged and young mice. Although PC causes an increase in membrane fluidity of lymphocytes from both young and old mice, and from humans, this effect on fluidity is not prevented by a protein kinase inhibitor, while PC's effect on IgD-R upregulation is prevented by the inhibitor. Moreover, no correlation was observed between IgD-R upregulation and membrane fluidity changes induced by AL721 administered in vivo. To evaluate the role of IgD-R induction in the augmentation of antibody production by phospholipids, the effect of monomeric IgD was investigated. The augmenting effect of AL721 on antibody production was prevented by a single injection of monomeric IgD at the time of antigen administration. We conclude that (1) PC-containing lipid mixtures are effective in enhancing antibody production in aged mice, (2) induction of IgD-R is responsible for the augmenting effects of AL721 on antibody production, and (3) monomeric IgD not only blocks the upregulation of IgD-R, as shown previously, but also the augmenting effect of previously upregulated IgD-R on T cells by preventing their interaction with surface IgD+ B cells.

Relationship between humoral immunoaugmenting properties of DHEAS and IgD-receptor expression in young and aged mice.

IgD-receptors are associated with augmented antibody production in vivo and are induced on CD4+ T cells by aggregated IgD in young but not in aged mice. In the current study orally or intraperitoneally administered DHEAS was found to enhance antibody production, as measured in a plaque-forming cell assay, and also to cause an increased expression of IgD-R on T cells in both young and aged mice. IgD-R+ T cells are enumerated by rosette cell formation with IgD-coated SRBC. Since, as shown previously, the immunoaugmenting effect of IgD-R+ T cells is blocked by injection of monomeric IgD, the effect of monomeric IgD was also examined in DHEAS-pretreated mice. The inhibitory effect obtained with monomeric IgD in these studies indicates that upregulation of IgD-R by DHEAS plays an important role in the immunoenhancing effect of this hormone. In addition, no significant effect of DHEAS was obtained on contact hypersensitivity to DNCB or on proliferative responses of T cells from young or aged mice. Aged but not young mice showed increases in the numbers of Ia+ epidermal Langerhans cells after DHEAS treatment.

Soluble beta 2-microglobulin-free class I heavy chains are released from the surface of activated and leukemia cells by a metalloprotease.

Regulation of the expression of major histocompatibility complex (MHC) class I heavy chains not associated with beta 2-microglobulin (beta 2m) on freshly isolated and in vitro cultured human B and T leukemia cells was analyzed. These beta 2m-free class I heavy chains originate from surface beta 2m-associated MHC class I molecules and are expressed as integral membrane glycoproteins on activated, but not resting, cells. We found that the levels of beta 2m-free class I heavy chains can be regulated by proteolytic cleavage and release into the medium of soluble molecules containing the extracellular domains. The release is mediated by a Zn(2+)-dependent, membrane-bound metalloprotease that does not cleave HLA-DR, CD4, and CD71 surface receptors and can be activated by phorbol myristate acetate. Specific cleavage by the metalloprotease occurs at a site close to the papain cleavage site in the alpha 3 domain of class I heavy chains. This site is not accessible to the metalloprotease in beta 2m-associated MHC class I molecules. The dissociation of beta 2m-associated MHC class I molecules and subsequent cleavage of beta 2m-free class I heavy chains may be partially responsible for controlling the levels of MHC class I molecules on the surface of activated cells.

Immunophenotypic evaluation of the bone marrow in non-Hodgkin's lymphoma.

Immunophenotypic evaluations of the bone marrow (BM) are reported on 69 aspirates from 58 patients who had non-Hodgkin's lymphoma or chronic lymphocytic leukemia involving the BM. Using flow cytometry and immunofluorescence microscopy on density gradient isolated BM mononuclear cells, the neoplasm could be identified and characterized in 59 aspirates (86%) from 49 patients (84%). Using International Working Formulation guidelines the neoplasms were classified on the basis of prior or subsequent histopathology of lymph node, spleen, skin, or other soft tissue site, or by evaluation of peripheral blood in chronic lymphocytic leukemia. In nine cases the lymphoma could not be completely classified according to International Working Formulation guidelines because only BM was available for evaluation. The neoplasm in the BM was identified and characterized immunophenotypically in all 29 cases of chronic lymphocytic leukemia/well-differentiated lymphocytic lymphoma (WDLL) (100%), in 11 of 12 cases of low-grade lymphoma other than WDLL (92%), in 11 of 15 cases of intermediate-grade lymphoma (73%), and in two of four cases of high-grade lymphoma (50%). Six of the nine cases not classified by International Working Formulation guidelines could be characterized immunophenotypically. In 10 cases immunophenotypic studies revealed negative findings, although the concurrent core biopsy specimens were positive. In two cases immunophenotypic studies with positive findings accompanied a negative core biopsy specimen. A panel of immunohistochemical reagents reactive with fixative/paraffin-resistant antigens was used for a retrospective evaluation of the 69 core biopsy specimens. When compared with the immunophenotypic data obtained from the marrow aspirates these results proved to be only moderately reliable in B-lineage neoplasms and unreliable in T-cell neoplasms. Thus, immunophenotyping of aspirated marrow by flow cytometry was found to be the most reliable method for determining the antigenic profiles of BM-based lymphomas.

Monoclonal gammopathy of undetermined significance: a morphologic and immunophenotypic study of the bone marrow.

A monoclonal paraprotein in the serum or urine raises the possibility of myeloma. However, in a significant proportion of individuals with serum paraproteins, particularly those with low levels of paraprotein, clinical and routine bone marrow evaluation is not diagnostic of an underlying neoplasm. The purpose of this study was to define the pathologic basis for monoclonal gammopathy in patients whose bone marrow biopsies showed no evidence of myeloma. We used immunofluorescence microscopy and flow cytometry of cell suspensions prepared from aspirated marrow, as well as immunohistochemistry of core biopsies, to perform immunopathologic evaluations of the bone marrow from 26 such patients. Eighteen patients with myeloma and seven without a serum paraprotein or evidence of myeloma were similarly studied. The data indicate that 17 of the 26 patients with monoclonal paraproteins whose routine bone marrow biopsies were normal or nondiagnostic had, in fact, a dispersed monotypic plasma cell population of concordant immunoglobulin heavy and light chain type in the bone marrow demonstrable by at least one of the three analytic methods. Among these, immunofluorescence microscopy of isolated bone marrow mononuclear cells was the most sensitive assay. Immunophenotypic evaluation of the bone marrow is useful for documenting and quantifying a monoclonal plasma cell population in patients with monoclonal gammopathy of undetermined significance.

IgM monoclonal gammopathy/Waldenström's macroglobulinemia: a morphological and immunophenotypic study of the bone marrow.

The presence of a monoclonal paraprotein in the serum or urine raises the possibility of myeloma or lymphoma/leukemia. Yet, in a significant proportion of individuals with serum paraproteins, particularly those with low levels of paraprotein, clinical and routine bone marrow evaluation is not diagnostic of an underlying neoplasm. The purpose of this study was to define the pathologic basis for macroglobulinemia in patients whose routine bone marrow biopsies were not diagnostic of a lymphoplasmacytic neoplasm. We used immunofluorescence microscopy and flow cytometry of cell suspensions prepared from aspirated marrow, as well as immunohistochemistry of core biopsies, to perform immunopathologic evaluations of the bone marrow from 16 such patients. Seven individuals without a monoclonal serum paraprotein, who were similarly studied, served as controls. Our data indicate that 13 of the 16 patients with monoclonal serum IgM paraproteins whose routine bone marrow biopsies were normal or showed nondiagnostic changes morphologically had a dispersed monotypic B lineage population of concordant immunoglobulin heavy and light chain type in the bone marrow. The immunophenotype of these cells spanned the range from mature B cell to plasmacytoid B cell to plasma cell. In four of these 13 patients a diagnosis of lymphoplasmacytic lymphoma could be made on the basis of greater than or equal to 20% monoclonal B lineage cells among bone marrow mononuclear cells.

New family of binary arrays for coded aperture imaging.

We introduce a new family of binary arrays for use in coded aperture imaging which are predicted to have properties and sensitivity (SNR) equal to that of the uniformly redundant array (URA). The new arrays, called MURAs (modified URAs), have decoding coefficients all of which are unimodular, resulting in a reconstructed image with noise terms completely independent of image-source structure. Although the new arrays are derived from quadratic residues, they do not belong to the cyclic difference set or set of pseudonoise sequences and consequently are constructible in configurations forbidden to those designs, thus providing the user with a wider selection of aperture patterns to match his particular needs. With the addition of MURAs to the family of binary arrays, all prime numbers can now be used for making optimal coded apertures, increasing the number of available square patterns by more than a factor of 3.

Proliferative and cytotoxic immune functions in aging mice. IV. Effects of suppressor cell populations from aged and young mice.

The changes with age in three splenic suppressor cell populations were studied in C57BL/6 mice. Allospecific Ts cells and nonspecific non-T suppressor cells were both generated in vitro in allogeneic MLC. The presence of "pre-existing" suppressor cells in fresh spleen cells from normal mice was examined. Suppressor cell activities were assayed for their ability to suppress proliferation in a fresh allogeneic MLC after treatment to prevent their own proliferation. The ability to generate both specific and nonspecific suppressor cells decreased with age, whereas pre-existing suppressor cells were detected in spleens from the majority of the aged animals but not in spleens from young animals. The decrease in suppressor cell activity was not due to any requirement for age matching between donors of suppressor and target cells. The specific and nonspecific MLC-generated suppressor cells inhibited both the proliferative response in the assay MLC and the generation of cytotoxic cells. The pre-existing suppressor cells only suppressed the proliferative response and not the generation of cytotoxic cells. The changes seen with age in these suppressor cell populations suggest that the ability to generate suppression (both allospecific and nonspecific) to newly encountered Ag declines with age, whereas a resident splenic suppressor cell population accumulates over the lifetime of the animals.

Physiology of IgD. VIII. Age-related decline in the capacity to generate T cells with receptors for IgD and partial reversal of the defect with IL 2.

In previous studies we have shown that T cells from young mice can be induced to express receptors for IgD (T delta cells) after in vivo or in vitro exposure to IgD or IL 2. In the present study, mice of three different strains were used to study the effects of aging on the ability of splenic T cells to express such receptors. It is shown that T cells from mice older than 18 mo of age are deficient with respect to their ability to express receptors for IgD after in vivo or in vitro exposure to IgD. Similarly, IFN-gamma induces T delta cells in young but not in aged mice. In contrast, T delta cells can be induced in aged mice with IL 2, although to a lower percentage than in young mice. In agreement with our previously reported findings, the current study demonstrates the immunoaugmenting effect of IgD injections on the antibody response in young mice. This effect is absent in aged mice and thus correlates with the failure of IgD to induce T delta cells in such animals.

Proliferative and cytotoxic immune functions in aging mice. III. Exogenous interleukin-2 rich supernatant only partially restores alloreactivity in vitro.

The ability of exogenous interleukin-2 (IL-2) rich supernatant to restore the defective T cell mediated immune functions of spleen cells from aged C57BL/6 mice was analyzed. Addition of IL-2 rich supernatant to allogeneic mixed lymphocyte cultures (MLC) resulted in an increase in the proliferative response of spleen cells from both young and old mice. The MLC response of cells from old mice was, however, not restored to the level of proliferation seen with splenocytes from young animals. In studying the generation of specific T cell suppressor function, it was found that IL-2 rich supernatant enhanced this function only for spleen cells from those aged animals which demonstrated a defective response in its absence. The response of these mice was thereby restored to the normal level. The response of cells from young control animals and aged mice with normal suppressor activity was not affected by the addition of IL-2 rich supernatant. We conclude that decreased IL-2 production constitutes a functionally important aspect, but is by no means the only defect in the immune response of aged mice. The results also suggest that responsiveness to IL-2 is less affected by age than lymphokine production.

Proliferative and cytotoxic immune functions in aging mice. II. Decreased generation of specific suppressor cells in alloreactive cultures.

The ability of spleen cells from aged C57BL/6 mice to generate specific suppressor cells in mixed lymphocyte cultures (MLC) against allogeneic H-2 antigens was investigated. The suppressor cells from young and old mice were assayed in parallel for their ability to inhibit the proliferative response and the generation of cytotoxicity in fresh MLC. Suppressor cell generation was found to be significantly decreased in 41% of aged mice (23 to 28 mo) as compared to young controls (3-8 mo). The suppressor cells were H-2-specific, radiation-resistant (1000 R), and Thy-1+; they did not function by lysing the fresh stimulators or responder cells, or by absorbing the interleukin 2 in the fresh cultures. Suppression required very small numbers of cells to be effective. It was concluded that the effect of aging was less marked on specific suppressor cell generation than on generation of cytotoxic T cells in the MLC. However, a third type of response studied, the proliferative response, was affected earliest and most severely.

Modification of age-related immune decline in mice dietarily restricted from or after midadulthood.

Although weaning-initiated dietary restriction of rodents is known to increase maximum survivorship and inhibit spontaneous late-life disease and immunologic aging, restriction begun in adulthood has been much less thoroughly evaluated. In the present studies, male mice of a long-lived F1 hybrid strain were gradually restricted dietarily beginning at 12 mo or older until their body weights stabilized at 60-70% of controls. Underfeeding decreased the number of nucleated cells per spleen but increased the percentage of T cells. For mice restricted at 12, 17, or 22 mo and tested at various ages thereafter, the [3H]thymidine uptake of spleen cells after phytohemagglutinin stimulation significantly exceeded values for age-matched unrestricted controls. Restriction did not, however, alter either splenocyte responses to concanavalin A or to B-cell mitogens or phytohemagglutinin responses of peripheral lymph node cells. In the splenic plaque-forming cell response to injected sheep erythrocytes, restricted and control mice differed more clearly in response kinetics than in peak levels. The splenic cell-mediated lymphocytotoxic response to alloantigens was comparable in old mice (27-29 mo) restricted since 12 mo of age with that of young (5- to 6-mo) controls and was greater than that of age-matched old controls. Spontaneous tumors were observed less frequently in 19- to 25-mo-old mice restricted at 12 mo of age than in mice restricted at 17 mo or in controls. Our results indicate that appropriate food restriction initiated in adulthood influences immunosenescence and spontaneous tumor incidence in a fashion not unlike its weaning-initiated counterpart.

Proliferative and cytotoxic immune functions in ageing mice. I. Sequence of decline of reactivities measured under optimal and suboptimal sensitization conditions.

Individual young adult, middle-aged, and old C57BL/6J male mice were tested for in vitro generated proliferative and cytotoxic responses to H-2 alloantigens under a variety of sensitization conditions. Proliferation in mixed lymphocyte culture (MLC) had decreased by 14 months of age (middle-aged), whether measured by directly assaying cultures in microtitre plates (micro MLC) or by labelling aliquots taken from large culture tubes (macro MLC). Cytotoxicity did not decline until a later age if sensitization was done in large tubes (macro cell-mediated lympholysis, CML). When cytotoxic activity was assayed by measuring lysis after addition of chromated cells to MLCs in microtitre plates (micro CML), differences were revealed between young and middle-aged animals. However, these conditions were suboptimal for generation of cytotoxicity even in young controls and showed even lower responses in the middle-aged group. It was concluded that proliferation showed an earlier, more severe decline than cytotoxicity with age as the proliferative response had declined by middle-age under all sensitization conditions used. With optimal sensitization conditions, senescent mice (26--30 months) showed a four- to ten-fold decrease in cytotoxicity compared with young adult mice.

Cellular immunity during pregnancy. II. Response to T and B cell mitogens.

Immune reactivity of primiparous pregnant C57Bl/6J mice was investigated using in vitro assays of mitogen reactivity. The response to the T cell mitogens phytohemagglutinin (PHA) and concanavalin A of cells from the paraaortic (PA) lymph nodes, which drain the uterus, was decreased in pregnant animals. Reactivity to lipopolysaccharide, a B cell mitogen, was normal. The decreased PHA response was seen with PA cells from mice bearing syngeneic or allogeneic (to DBA/2J) fetuses. It was not due to a change in sensitivity to PHA dose or to active suppression (as demonstrated by mixing experiments). Phytohemagglutinin reactivity of cells from inguinal nodes of pregnant mice showed a more variable depression of response in comparison to that seen with cells from the draining PA nodes. The response of axillary and brachial node cells was similar to virgin values. Statistical analysis revealed no differences in the average number of PA lymphocytes or fetuses per mouse between mice bearing syngeneic or allogeneic fetuses. This parallels the similarities found between syngeneic and allogeneic matings in in vitro functional assays. This study demonstrates that pregnant mice (syngeneic or allogeneic) show only a decrease in T proliferative capacity localized to the area of the uterus, while such responses in the rest of the body are left essentially intact.

Cellular immunity during pregnancy. I. Proliferative and cytotoxic reactivity of paraaortic lymph nodes.

The possibility of changes in immune reactivity during pregnancy was studied by measuring cellular immunity in vitro of the paraaortic (PA) lymph nodes, which drain the uterus in pregnant mice. The proliferation of PA lymph node cells from primiparous pregnant C57Bl/6J mice, in mixed lymphocyte cultures (MLC) against alloantigens, was lower in magnitude, but had the same kinetics, as the response of virgins. This was observed in syngeneic and allogeneic (to DBA/2J) pregnancies, and using the paternal as well as third party allogeneic stimulators. The response was depressed by day 8 of gestation and returned to normal two days after delivery. The decrease was not due to an active suppressor mechanism, as assayed by mixing experiments. Irradiation (1500R) of the lymphocytes from pregnant mice, prior to mixing with cells from virgins, did not reveal the presence of a radioresistant suppressor cell. In contrast to the MLC results, no differences were found between lymphocytes from pregnant and virgin mice in their ability to develop MLC-generated cell-mediated lympholysis (CML) against alloantigens, as measured in a chromium release assay. The PA lymph node cells from pregnant animals bearing allogeneic fetuses also did not show evidence of in vivo sensitization to the paternal alloantigens. Therefore, the local nodes draining the uterus from primiparous pregnant animals bearing syngeneic or allogeneic fetuses show a nonspecific decrease of MLC proliferation while retaining the capacity to generate normal CML activity.