How to Diagnose and Treat CD5-Positive Lymphomas Involving the Spleen.

Patients with CD5-expressing lymphomas presenting with splenomegaly are frequently diagnosed with chronic lymphocytic leukemia. The most important differential diagnosis is mantle cell lymphoma, both in its classical and leukemic, non-nodal forms, given its prognostic and therapeutic implications. Other small B-cell neoplasms that frequently involve the spleen and occasionally express CD5 include the splenic marginal zone lymphoma, hairy cell leukemia and, rarely, lymphoplasmacytic lymphoma. The frequency of CD5 positivity depends in part on the sensitivity of the detection methods employed. Usually, a combination of morphological, immunophenotypic and molecular findings allows for a precise sub-classification of CD5-positive, low-grade B-cell lymphomas of the spleen. Some of these tumors may display a mixture of small and larger B cells, raising the possibility of more aggressive lymphomas, such as diffuse large B-cell lymphomas (DLBCL). Approximately 5-10% of DLBCL are CD5-positive and some may manifest as primary splenic lesions. When available, the morphology of DLBCL in the splenic tissue is distinctive and a leukemic picture is very rare. In conclusion, the appropriate morphological and clinical context assisted by flow cytometry panels and/or immunohistochemistry allows the differential diagnosis of CD5-positive, non-Hodgkin, B-cell lymphomas involving the spleen.

Low-Grade Primary Splenic CD10-Positive Small B-Cell Lymphoma/Follicular Lymphoma.

Primary splenic lymphoma (PSL) is a rare malignancy representing about 1% of all lymphoproliferative disorders, when using a strict definition that allows only involvement of spleen and hilar lymph nodes. In contrast, secondary low-grade B-cell lymphomas in the spleen, such as follicular lymphomas (FL), lymphoplasmacytic lymphoma and chronic lymphocytic leukemia/ small lymphocytic lymphoma, particularly as part of advanced stage disease, are more common. Indolent B cell lymphomas expressing CD10 almost always represent FL, which in its primary splenic form is the focus of this review. Primary splenic follicular lymphoma (PSFL) is exceedingly infrequent. This type of lymphoproliferative disorder is understudied and, in most cases, clinically characterized by splenomegaly or cytopenias related to hypersplenism. The diagnosis requires correlation of histopathology of spleen, blood and/or bone marrow with the correct immunophenotype (determined by flow cytometry and/or immunohistochemistry) and if necessary, additional molecular profiling. Management of this incurable disease is evolving, and splenectomy remains the mainstream treatment for stage I PSFL.

Differences amid bone marrow and cord blood hematopoietic stem/progenitor cell division kinetics.

Human hematopoietic stem/progenitor cells (HSC) isolated based upon specific patterns of CD34 and CD38 expression, despite phenotypically identical, were found to be functionally heterogeneous, raising the possibility that reversible expression of these antigens may occur during cellular activation and/or proliferation. In these studies, we combined PKH67 tracking with CD34/CD38 immunostaining to compare cell division kinetics between human bone marrow (BM) and cord blood (CB)-derived HSC expanded in a serum-free/stromal-based system for 14 days (d), and correlated CD34 and CD38 expression with the cell divisional history. CB cells began dividing 24 h earlier than BM cells, and significantly higher numbers underwent mitosis during the time in culture. By d10, over 55% of the CB-cells reached the ninth generation, whereas BM-cells were mostly distributed between the fifth and seventh generation. By d14, all CB cells had undergone multiple cell divisions, while 0.7-3.8% of BM CD34(+) cells remained quiescent. Furthermore, the percentage of BM cells expressing CD34 decreased from 60.8 +/- 6.3% to 30.6 +/- 6.7% prior to initiating division, suggesting that downmodulation of this antigen occurred before commencement of proliferation. Moreover, with BM, all primitive CD34(+)CD38(-) cells present at the end of culture arose from proliferating CD34(+)CD38(+) cells that downregulated CD38 expression, while in CB, a CD34(+)CD38(-) population was maintained throughout culture. These studies show that BM and CB cells differ significantly in cell division kinetics and expression of CD34 and CD38, and that the inherent modulation of these antigens during ex vivo expansion may lead to erroneous quantification of the stem cell content of the expanded graft.

Mobilization strategies for the collection of peripheral blood progenitor cells: Results from a pilot study of delayed addition G-CSF following chemotherapy and review of the literature.

OBJECTIVE: Given the potential to limit cost, we conducted a pilot study evaluating delayed, low-dose granulocyte colony-stimulating factor (G-CSF) following chemotherapy for the procurement of peripheral blood progenitor cells (PBPCs) for autologous transplantation and reviewed the relevant literature. PATIENTS AND METHODS: Twenty-eight patients with various malignancies received cyclophosphamide 4 gm/m(2) and paclitaxel 170 mg/m2 followed by G-CSF 300 microg/d or 480 microg/d starting day +5 until two to four daily large volume leukapheresis yielded > or =5.0 x 10(6) CD34+ cells. We searched MEDLINE, Pubmed, and EMBASE databases from 1990 to the present to identify papers on PBPC procurement using delayed G-CSF (starting day +4 or later) following chemotherapy. RESULTS: G-CSF was administered for a median of 9 days at an average cost of 1260 USD per 70-kg patient. Collection was initiated at a median of 12 days after chemotherapy. A median 2.5 (range 2-4) apheresis were performed yielding an average daily CD34+ collection of 6.9 x 10(6)/kg (range 0.35-56.7). After one apheresis, 82% and 57% of patients collected > or =2.5 x 10(6)/kg and > or =5.0 x 10(6)/kg, respectively. Ultimately, 89% collected > or =5.0 x 10(6)/kg. Febrile neutropenia and catheter-related infection developed in five and two patients, respectively. All patients proceeded to transplantation and engrafted successfully with a median of 14.9 x 10(6)/kg (range 1.05-113) cells infused. Eleven published reports were identified involving 590 patients of whom 498 received G-CSF at a dose range of 250 microg/d to 10 microg/kg/d starting day +4 to 15 for a period of 4 to 9 days for PBPC procurement. Among these reports, 62 to 100% and 33 to 96% of patients collected > or =2 to 2.5 x 10(6) and > or =5.0 x 10(6) CD34+ cells, respectively. CONCLUSION: The use of delayed, low-dose G-CSF plus chemotherapy for stem cell mobilization was feasible and provides further evidence supporting this potentially cost-effective strategy. A review of the literature supports our findings and emphasizes the need for larger studies to address this issue.

IL-15-mediated induction of LFA-1 is a late step required for cytotoxic differentiation of human NK cells from CD34+Lin- bone marrow cells.

Optimal differentiation of cytotoxic NK cells is important to provide protective innate immunity to patients after bone marrow transplantation. In vitro differentiation of CD56(+)CD3(-) NK cells takes weeks and is supported by several cytokines, including IL-2, IL-7, and IL-15, and thus can be useful for immunotherapy. However, IL-2 therapy is problematic in vivo, and NK cells differentiated in vitro with only IL-7 lack cytotoxicity. We assessed whether human NK cells initially differentiated in vitro from CD34(+)Lin(-) bone marrow cells with IL-7 could acquire cytotoxicity after exposure to additional cytokines and what changes promoted cytotoxicity. The cells cultured with IL-7 already had granzyme B as well as perforin, as previously reported, the proteins of cytotoxic granules. The cells also lacked LFA-1. After 1 wk of secondary culture with either IL-2 or IL-15, but not with IL-12 or IL-18, the IL-7-cultured cells acquired cytotoxicity. IL-2 or IL-15 also induced LFA-1. Ab to the LFA-1 subunits CD11a and CD18 blocked lysis by the NK cells, indicating that the new LFA-1 correlated with, and was essential for, the cytotoxic function of the in vitro generated cells. The LFA-1 also participated in target cell binding by the in vitro differentiated cells. In this study, we demonstrated a new function for IL-15, the induction of LFA-1 in NK progenitor cells, and that IL-15 does more than merely support NK progenitor cell proliferation. The efficacy after only 1 wk of IL-15 administration is a positive practical feature that may apply to human therapy.

Human natural killer cell development in a xenogeneic culture system.

In vivo and in vitro xenogeneic models have shown the ability of a non-human environment in supporting human haemopoiesis. In the present study, we evaluated the effect of fetal sheep thymic stroma in the in vitro development of natural killer (NK) cells from human haemopoietic progenitors. CD34+HLA-DR+ (CD34+ DR+)Lin- and CD34+DR-Lin- bone marrow (BM) progenitors were cultured for 3 weeks with or without interleukin 2 (IL-2), in fetal sheep thymic stroma contact and transwell cultures. Both progenitors gave rise to NK cells, defined as CD45+CD56+ cells, in the presence or absence of IL-2; however, the percentage of NK cells originated in cultures with IL-2 was significantly higher. Direct contact with stroma seemed to be required for the most immature progenitors, CD34+DR-Lin-, to differentiate along the NK cell lineage. Functional assays revealed that only cells grown in the presence of IL-2 were cytolytic against K562 targets and, curiously, NK cells derived from CD34+DR-Lin- progenitors were more cytotoxic that NK cells derived from CD34+DR+Lin- progenitors. These studies suggest that the ability of fetal sheep thymic stroma in promoting the generation of human NK cells from haemopoietic progenitors may have relevance in terms of NK cell ontogeny and induction of tolerance in transplantation.