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Prepublished online as a Blood First Edition Paper on October 3, 2002; DOI 10.1182/blood-2002-02-0469.
IMMUNOBIOLOGY
From the Department of Research and Development, EFS
Rhône-Alpes, and Research Group on Lymphoma, Albert Bonniot
Institute, La Tronche, France; and Clinical Hematological Department,
Michallon Hospital, Grenoble, France.
To assess the sensitivity of primary non-Hodgkin lymphoma cells to
rituximab-mediated cytotoxicity, we compared the potency of several
rituximab-mediated killing mechanisms on fresh lymphoma cells. All
lymphoma cells tested were equally sensitive to antibody-dependent cell-mediated cytotoxicity (ADCC), antibody-mediated phagocytosis of
tumor cells, and rituximab-induced apoptosis. However, they were
differentially lysed by complement-dependent cytotoxicity (CDC). We found that taking into account both CD20 and complement regulatory protein expression on tumor cells could predict CDC sensitivity in vitro. Importantly, the sensitivity of lymphoma cells to
CDC was consistent with the reported different clinical response rates
of lymphomas: rituximab induced high CDC killing of follicular
lymphoma cells, whereas mantle cell lymphoma and diffuse large cell lymphoma cells were moderately sensible to CDC, and small lymphocytic lymphoma cells were almost all
resistant. We propose that CDC is a determinant mechanism of
rituximab-induced killing in vivo. Poor sensitivity to CDC in vitro
might predict a poor clinical response, whereas high sensitivity to CDC
would only indicate a likelihood of response to rituximab treatment.
(Blood. 2003;101:949-954) Rituximab is a chimeric mouse/human antibody,
bearing the human IgG1 and Cells
Antibodies and flow cytometry
Three-color flow cytometry phenotyping of tumor cells was performed on a FACScan flow cytometer (Becton Dickinson, Mountain View, CA). Saturating amounts of antibodies were added to cells for 30 minutes at 4°C, before extensive washing and flow cytometry analysis. Mean fluorescence intensity (MFI) values were used as a semiquantitative measure of the expression of CD20 and of complement inhibitors. All phenotypes were performed on the same day as CDC assays. CD20-induced apoptosis of tumor cells Tumor cells (1 × 106/mL) were incubated in complete medium supplemented by 10% heat-inactivated human serum (obtained from voluntary donors at the Etablissement Français du Sang, La Tronche, France), in the presence or absence of 2 µg/mL rituximab. Apoptosis of cells was followed at day 0, day 1, and day 2 by staining with 2.5 µg/mL propidium iodide (PI; Immunotech) and flow cytometer analysis. Annexin V staining of apoptotic cells could not be reliably measured because rituximab induced cell clusters with ambiguous higher annexin V staining. PI staining was correlated to DiOC6(3) (Molecular Probes, Eugene, OR) or the TUNEL assay (Mebstain, Immunotech), but not to annexin V staining (see "Results and Discussion"). In preliminary experiments, graded doses of soluble rituximab (0, 0.02, 0.2, 2, 20 µg/mL) were added to the cells in test tubes, with or without cross-linking of rituximab by antihuman IgG goat antibody F(ab')2 (Immunotech) or affinity-purified goat antihuman antibodies (Biosys, Compiègne, France) at 2 or 20 µg/mL, or rituximab was first immobilized on microplates before adding the cells. No significant effect of cross-linking was observed, and the optimal condition for apoptosis induction was chosen as 2 µg/mL soluble rituximab. Because of the high rate of spontaneous apoptosis of some tumor cells, CD20-induced apoptosis could not be determined as the percentage of remaining viable cells with anti-CD20 as compared to cells without antibody at the same time. It was therefore calculated according to the following formula: 100 × (% viable cells at day 1 without antibody % viable cells with
rituximab at day 1)/(% viable cells at day 0). Human pooled
immunoglobulins (Tégéline, LFB, Les Ulis, France;
containing 25% IgG1 antibodies and used as an isotype control for
rituximab) had no effect at any concentration tested (not shown).
Phagocytosis Tumor cells were opsonized for 30 minutes at 4°C by 2 µg/mL rituximab and then washed. Macrophages were generated as described11 and were added to tumor cells (1 macrophage for 5 tumor cells) in RPMI 10% heat-inactivated human serum. After 2 hours, cells were cytospun and stained for visual counting of phagocytosis. Results are presented as the percentage of phagocytosing macrophages (macrophages that have phagocytosed at least one tumor cell). For inhibition experiments, macrophages were incubated with 5 µg/mL CD16 (BD Pharmingen, Heidelberg, Germany), CD32 (Stem Cell Technologies, Meylan, France), or CD64 (Diaclone Research, Besançon, France) or their isotype control as blocking antibodies during phagocytosis.ADCC Monocytes and natural killer (NK) cells were purified from freshly collected blood using Rosette Sep isolation kits (Stem Cell Technologies). Polynuclear neutrophils were obtained by blood sedimentation on dextran and Ficoll-Hypaque density gradient centrifugation. Purity of effectors was determined by flow cytometry analysis and was 85% for NK cells, 70% for monocytes, 99% for neutrophils (based on forward and side scatter [FSC-SSC] profiles, CD14, CD16, and CD56 expression).The cytotoxicity of effector cells in the presence or absence of 2 µg/mL rituximab was measured in a standard 4-hour
51Cr-release assay, as previously described.12
Briefly, 104 51Cr-labeled tumor cells were
mixed with the effector cells at different effector-target
(E/T) ratios (25:1-0.01) in RPMI 10% heat-inactivated human serum.
After a 4-hour incubation at 37°C in 5% CO2 in air, the
radioactivity in the supernatants was counted. The percentage of
specific lysis was calculated according to the following formula: % lysis = 100 × (ER Complement-mediated lysis Rituximab was added to tumor cells (1 × 106 /mL) in complete medium supplemented by human serum, inactivated or not by incubation at 56°C for 30 minutes. After 2 hours at 37°C, cell lysis was determined by PI staining of cells and analysis by flow cytometry. Lysis was calculated according to the following formula: 100 × (% viable cells with inactivated serum % viable
cells with native serum)/(% viable cells with inactivated serum), all
measures being taken after the 2-hour incubation.
Statistical analysis All statistical analyses were performed using Statview software (Abacus Concepts, Berkeley, CA).
In this paper, we examined cell autonomous factors that could account for rituximab efficacy. This was warranted by the observation that different lymphoma histologic types have different clinical outcomes, suggesting tumor-intrinsic influencing factors on rituximab efficacy. We therefore measured in vitro the intrinsic sensitivity of primary lymphoma cells to different mechanisms of antibody-mediated cytotoxicity. We first measured expression of CD20 molecules using a semiquantitative
method on 28 tumor samples from distinct histologic types (7 FLs, 7 MCLs, 7 SLLs, and 7 DLCLs) and on 3 NT B-cell suspensions. As expected,
Figure 1 shows that all tumor cells expressed CD20 antigen but with distinct intensities, depending on
lymphoma groups. SLL cells expressed the lowest level of CD20, and DLCL
cells the highest. CD20 expression was found at an intermediate level
in FL cells and MCL cells.
We then tested the rituximab-induced apoptosis for all tumor and NT
samples. Several reports on lymphoma cell lines have shown a direct
effect of rituximab in apoptosis induction, documented at the molecular
level,5,13,14 and which is currently viewed as a major
mechanism of rituximab efficacy.15 However,
rituximab-induced apoptosis has not been extensively tested on primary
lymphoma cells, and its importance remains to be evaluated. To
determine the magnitude of CD20-induced apoptosis on fresh malignant
cells, rituximab was added to lymphoma cells, and cell viability was followed by flow cytometry after 1 or 2 days. We used different techniques to analyze rituximab-induced cell death: annexin V staining
to detect exposed phosphatidylserine, TUNEL method to reveal the 3'-OH
DNA ends, DiOC6(3) labeling to monitor mitochondrial transmembrane potential disruption, and finally PI to allow
discrimination of viable from nonviable cells. As illustrated in Figure
2 for patient FL3, we found that
rituximab induced annexin V staining, often accompanied by clustering
of cells (cells with increased FSC), which could not be disrupted.
However, in concordance with published results,16
the significance of annexin V staining was not clear. Indeed, there was
a good correlation between PI staining, TUNEL, and DiOC6(3)
at day 2, but no correlation with annexin V staining. This indicates
that annexin V cannot reliably be taken as a marker for
rituximab-induced apoptosis, and this is why we used PI exclusion as a
viability marker. Spontaneous apoptosis in absence of rituximab was
highly variable during the 2-day observation period for the 28 tumor
samples (between 0% and 77% at day 1; Table
1), impairing accurate evaluation of apoptosis at later time points. Rituximab did induce apoptosis of
lymphoma cells, but, in the majority of cases, this effect was modest;
a mean of 10% CD20-induced apoptosis was found after 24 hours (Table
1) and about 15% at 48 hours when evaluable (data not shown). However,
this apoptosis was specific to CD20 because irrelevant immunoglobulins
had no effect (data not shown). Interestingly, NT B cells were not
sensitive to rituximab-induced apoptosis. In our hands,
cross-linking of rituximab, either by plate coating or by addition of
secondary antibodies, did not significantly alter apoptosis induction
(data not shown). Therefore, primary lymphoma cells are weakly
sensitive to rituximab-induced apoptosis. Moreover, no significant
differences were observed between FL, MCL, SLL, and DLCL (Table 1),
which questions its importance in vivo when considering the
differential clinical responses of lymphomas.
Because tumor cells did not appear to be intrinsically very sensitive
to a direct effect of rituximab, we examined the involvement of
cellular effectors that may be recruited to the tumor site. Tumor cell
clearance by antibody-dependent phagocytosis of whole lymphoma cells
was measured with macrophages as tissue cells that could infiltrate
tumor lesions.17 We generated macrophages from blood
monocytes11 and tested their capacity to phagocytose
rituximab-opsonized lymphoma cells in the presence of human serum. As
shown in Figure 3A, macrophages could
engulf rituximab-opsonized tumor cells (4 FLs, 4 MCLs, 4 DLCLs, 4 SLLs,
and 2 NTs), whereas tumor cells were only marginally internalized if
not opsonized. In this assay, SLL cells were less readily phagocytosed,
possibly due to their lower expression of CD20 (Figure 1). Inhibition
experiments with blocking antibodies indicated that the low-affinity
IgG receptor CD32/Fc
We then tested ADCC by different Fc
Therefore, primary lymphoma cells can be cleared by cellular effectors in the presence of rituximab, either by phagocytosis or by ADCC, which are likely to be important in vivo.19 However, the magnitude of effector cell recruitment after rituximab infusion is unknown, and, as for rituximab-induced apoptosis, lymphoma cells were equally susceptible to rituximab-mediated phagocytosis or ADCC, raising doubts about the in vivo relevance of these mechanisms to explain the different response rates of lymphomas. Besides cellular cytotoxicity, a potent mechanism of antibody-dependent
tumor cell killing is complement-mediated lysis.8 Human
cells are normally protected against spontaneous cytolytic activity of
complement by a battery of regulatory proteins, present in a soluble
form in the serum, or expressed on the cell surface.21 However, in the presence of rituximab linked to the tumor cell, activation of the classical pathway may overrun these regulatory mechanisms and lead to cell lysis. Therefore, expression of these inhibitory proteins by the tumor cells may be predictive of the outcome
of complement activation by rituximab, either death or survival.6,21 As preliminary studies, we performed kinetic and dose-response experiments, with cells from 2 patients, FL3 and FL4
(Figure 5). In the presence of rituximab,
CDC (abrogated after heat-inactivation of serum) was very efficient in
killing tumor cells, because up to 90% of cells were found to
incorporate PI after only 2 hours at 37°C, in optimal conditions.
These 2 patients displayed an identical expression of CD20; however,
they did not have the same sensitivity to CDC. Indeed, rituximab
induced dose-dependent killing of both cells, but cells from patient
FL3 were killed in the presence of 10% serum, whereas cells from
patient FL4 were not (Figure 5). Therefore, there seem to be other
factors regulating complement-dependent cell death, the effect of which depends on rituximab and serum concentration. To test the implication of complement regulatory proteins (CRPs) on tumor cells, we used a
semiquantitative method to determine CRP expression on cells from a
number of patients (Figure 6), and CDC
was measured in the presence of a fixed concentration of serum and
rituximab. To appreciate regulatory mechanisms, we chose nonsaturating
concentrations of rituximab (2 µg/mL) with sufficient serum
concentration (30%) and we analyzed CDC sensitivity in a 2-hour assay
of a panel of lymphoma cells (Figure 7).
Complement-dependent lysis ranged from 0% to 90% for the cells
tested. Cells that were resistant at 2 hours were still resistant at
further incubation times (data not shown). Cells were classified
according to their histologic type and it appeared that all FLs showed
some sensitivity to CDC (> 20% lysis, ranging from 20% to 90%;
Figure 7A). In contrast with a previous study,22 we found
that SLLs were almost all resistant, with 25% lysis in only one case
(Figure 7B), and MCLs were resistant except for 3 cases (lysis
We then attempted to correlate CDC sensitivity to the expression of CD20 and of the CRPs CD46, CD55, and CD59. First, SLLs expressed very low levels of CD20 (Figure 1), which could explain on its own their resistance to CDC (Figure 7B). However, when comparing FLs and MCLs, both expressed high amounts of CD20, but FLs were sensitive to CDC, whereas MCLs were resistant (Figures 1 and 7A,C). Globally, there was no direct correlation between lysis and expression of CD20 nor between lysis and expression of any CRP (data not shown). However, because CRPs may act synergistically to control complement activation (due to amplification loops in the complement activation cascade), we plotted the product of MFI values of CRP (INH; defined as the product of MFI values for CD46, CD55, CD59) against CD20 expression and CDC (Figure 7). This allowed distinguishing easily between FLs and MCLs; both substantially express CD20, but MCLs display high INH values, whereas FL had low INH values. There was no correlation between lysis and INH values (data not shown). On the other hand, cells expressing low levels of CD20 were poorly lysed (NT and SLL). This suggested that one might take the ratio of CD20/INH as an indication of sensitivity to CDC, and there was a significant correlation between the order of magnitude of CD20/INH and CDC (r = 0,8; P < .0001, t test; Figure 7F). Multivariate analysis using CD20, CD46, CD55, and CD59 expression as regressors gave a very similar regression (P < .0001 and r = 0.816). It has been suggested previously that CDC was directly correlated to CD20 expression, hence irrespective of CRP expression.22,23 However, this conclusion seems paradoxical with the claim that CDC is regulated by CRP, which is evidenced by the use of blocking antibodies,6,22 and is supported by this current study. We suggest that reported direct correlation between CD20 expression and CDC sensitivity rely on the saturating conditions used that do not allow regulation mechanisms to be evidenced. Based solely on CD20 expression, it is difficult to discriminate FL, MCL and DLCL cells. With the restriction that any correlation found in vitro depends on the experimental conditions used, we define a combination of 4 tumor-specific parameters as determining CDC lysis of lymphoma cells. However, augmenting rituximab dosing might induce better lysis of tumor cells, due to saturation of CD20 sites and overrunning of regulatory mechanisms (Figure 5), which could be translated as a therapeutic strategy.23-26 Our approach aimed at finding tumor cell-specific factors influencing rituximab killing in vitro. No striking differences were observed between primary lymphoma cells with regard to their sensitivity to rituximab-induced apoptosis, ADCC, and phagocytosis. However, CDC induced very different killing of tumor cells, with a pattern of sensitivity consistent with clinical data. In clinical studies, patients with FL display the best response rate to rituximab,3,4 whereas patients with MCL and DLCL have a moderate response,27 and SLL is associated with a poor or no clinical response to rituximab.28 Given that the other potential mechanisms of rituximab studied here (apoptosis, ADCC, phagocytosis) could not easily discriminate lymphoma cells, it is tempting to correlate in vitro CDC sensitivity to the probability of response according to the lymphoma histologic type; that is, poor sensitivity to in vitro CDC might predict a poor clinical response to rituximab. In the case of cells killed by CDC in vitro, this does not preclude the involvement of other factors finally determining the clinical efficacy of rituximab, and, indeed, a recent study found no correlation between sensitivity to CDC in vitro and clinical outcome within the FL histologic group.29 It is likely that supracellular factors, like tumor burden, or tumor vascularization are important parameters in determining rituximab efficacy. Moreover, complement activation is coupled to release of inflammatory chemotactic factors,30-33 recruiting cellular effectors for amplification and complexing of the antitumor response. In this perspective, CDC activation could be a necessary initial step (not necessarily massive) for recruitment of cells that may be critically involved, which is also consistent with its short kinetics. Intriguing clinical data are the occasional lag between rituximab infusion and therapeutic effect,34 which cannot easily be reconciled with immediate killing in short-term in vitro assays, but may indicate the involvement of late effectors. In line with this reasoning, rituximab-induced cross-presentation of tumor-derived antigens by dendritic cells was recently reported.35
Submitted February 20, 2002; accepted August 12, 2002.
Prepublished online as Blood First Edition Paper, October 3, 2002; DOI 10.1182/blood-2002-02-0469.
The publication costs of this article were defrayed in part by page charge payment. Therefore, and solely to indicate this fact, this article is hereby marked "advertisement" in accordance with 18 U.S.C. section 1734.
Reprints: Joël Plumas, Etablissement Francais du Sang-Grenoble, 29, Ave du Maquis du Gresivaudan, La Tronche Cedex 38701, France; e-mail: joel.plumas{at}wanadoo.fr and joel.plumas{at}efs.sante.fr.
1.
Reff ME, Carner K, Chambers KS, et al.
Depletion of B cells in vivo by a chimeric mouse human monoclonal antibody to CD20.
Blood.
1994;83:435-445
2.
Anderson KC, Bates MP, Slaughenhoupt BL, Pinkus GS, Schlossman SF, Nadler LM.
Expression of human B cell-associated antigens on leukemias and lymphomas: a model of human B cell differentiation.
Blood.
1984;63:1424-1433 3. McLaughlin P, Grillo-Lopez AJ, Link BK, et al. Rituximab chimeric anti-CD20 monoclonal antibody therapy for relapsed indolent lymphoma: half of patients respond to a four-dose treatment program. J Clin Oncol. 1998;16:2825-2833[Abstract].
4.
Maloney DG, Grillo-Lopez AJ, White CA, et al.
IDEC-C2B8 (rituximab) anti-CD20 monoclonal antibody therapy in patients with relapsed low-grade non-Hodgkin's lymphoma.
Blood.
1997;90:2188-2195
5.
Shan D, Ledbetter JA, Press OW.
Apoptosis of malignant human B cells by ligation of CD20 with monoclonal antibodies.
Blood.
1998;91:1644-1652
6.
Golay J, Zaffaroni L, Vaccari T, et al.
Biologic response of B lymphoma cells to anti-CD20 monoclonal antibody rituximab in vitro: CD55 and CD59 regulate complement-mediated cell lysis.
Blood.
2000;95:3900-3908 7. Clynes RA, Towers TL, Presta LG, Ravetch JV. Inhibitory Fc receptors modulate in vivo cytoxicity against tumor targets. Nat Med. 2000;6:443-446[CrossRef][Medline] [Order article via Infotrieve]. 8. Harjunpaa A, Junnikkala S, Meri S. Rituximab (anti-CD20) therapy of B-cell lymphomas: direct complement killing is superior to cellular effector mechanisms. Scand J Immunol. 2000;51:634-641[CrossRef][Medline] [Order article via Infotrieve]. 9. Plumas J, Chaperot L, Jacob MC, et al. Malignant B lymphocytes from non-Hodgkin's lymphoma induce allogeneic proliferative and cytotoxic T cell responses in primary mixed lymphocyte cultures: an important role of co-stimulatory molecules CD80 (B7-1) and CD86 (B7-2) in stimulation by tumor cells. Eur J Immunol. 1995;25:3332-3341[Medline] [Order article via Infotrieve]. 10. Sander CA, Kind P, Kaudewitz P, Raffeld M, Jaffe ES. The Revised European-American Classification of Lymphoid Neoplasms (REAL): a new perspective for the classification of cutaneous lymphomas. J Cutan Pathol. 1997;24:329-341[CrossRef][Medline] [Order article via Infotrieve]. 11. Chaperot L, Chokri M, Jacob MC, et al. Differentiation of antigen-presenting cells (dendritic cells and macrophages) for therapeutic application in patients with lymphoma. Leukemia. 2000;14:1667-1677[CrossRef][Medline] [Order article via Infotrieve].
12.
Plumas J, Jacob MC, Chaperot L, Molens JP, Sotto JJ, Bensa JC.
Tumor B cells from non-Hodgkin's lymphoma are resistant to CD95 (Fas/Apo-1)-mediated apoptosis.
Blood.
1998;91:2875-2885
13.
Mathas S, Rickers A, Bommert K, Dorken B, Mapara MY.
Anti-CD20- and B-cell receptor-mediated apoptosis: evidence for shared intracellular signaling pathways.
Cancer Res.
2000;60:7170-7176 14. Shan D, Ledbetter JA, Press OW. Signaling events involved in anti-CD20-induced apoptosis of malignant human B cells. Cancer Immunol Immunother. 2000;48:673-683[CrossRef][Medline] [Order article via Infotrieve]. 15. Cragg MS, French RR, Glennie MJ. Signaling antibodies in cancer therapy. Curr Opin Immunol. 1999;11:541-547[CrossRef][Medline] [Order article via Infotrieve]. 16. Weng WK, Levy R. Rituximab-induced annexin-V staining of tumor cells does not predict clinical outcome following rituximab treatment in follicular non-Hodgkin's lymphoma [abstract]. Blood. 2001;98:338aAbstract 1430. 17. Mantovani A, Bottazzi B, Colotta F, Sozzani S, Ruco L. The origin and function of tumor-associated macrophages. Immunol Today. 1992;13:265-270[CrossRef][Medline] [Order article via Infotrieve].
18.
Cartron G, Dacheux L, Salles G, et al.
Therapeutic activity of humanized anti-CD20 monoclonal antibody and polymorphism in IgG Fc receptor Fc
19.
Valerius T, Repp R, de Wit TP, et al.
Involvement of the high-affinity receptor for IgG (Fc gamma RI; CD64) in enhanced tumor cell cytotoxicity of neutrophils during granulocyte colony-stimulating factor therapy.
Blood.
1993;82:931-939 20. Abdullah N, Greenman J, Pimenidou A, Topping KP, Monson JR. The role of monocytes and natural killer cells in mediating antibody-dependent lysis of colorectal tumour cells. Cancer Immunol Immunother. 1999;48:517-524[CrossRef][Medline] [Order article via Infotrieve]. 21. Gorter A, Meri S. Immune evasion of tumor cells using membrane-bound complement regulatory proteins. Immunol Today. 1999;20:576-582[CrossRef][Medline] [Order article via Infotrieve].
22.
Golay J, Lazzari M, Facchinetti V, et al.
CD20 levels determine the in vitro susceptibility to rituximab and complement of B-cell chronic lymphocytic leukemia: further regulation by CD55 and CD59.
Blood.
2001;98:3383-3389
23.
Bellosillo B, Villamor N, Lopez-Guillermo A, et al.
Complement-mediated cell death induced by rituximab in B-cell lymphoproliferative disorders is mediated in vitro by a caspase-independent mechanism involving the generation of reactive oxygen species.
Blood.
2001;98:2771-2777
24.
O'Brien SM, Kantarjian H, Thomas DA, et al.
Rituximab dose-escalation trial in chronic lymphocytic leukemia.
J Clin Oncol.
2001;19:2165-2170 25. Keating M, O'Brien S. High-dose rituximab therapy in chronic lymphocytic leukemia. Semin Oncol. 2000;27:86-90[Medline] [Order article via Infotrieve].
26.
Byrd JC, Murphy T, Howard RS, et al.
Rituximab using a thrice weekly dosing schedule in B-cell chronic lymphocytic leukemia and small lymphocytic lymphoma demonstrates clinical activity and acceptable toxicity.
J Clin Oncol.
2001;19:2153-2164
27.
Coiffier B, Haioun C, Ketterer N, et al.
Rituximab (anti-CD20 monoclonal antibody) for the treatment of patients with relapsing or refractory aggressive lymphoma: a multicenter phase II study.
Blood.
1998;92:1927-1932
28.
Foran JM, Rohatiner AZ, Cunningham D, et al.
European phase II study of rituximab (chimeric anti-CD20 monoclonal antibody) for patients with newly diagnosed mantle-cell lymphoma and previously treated mantle-cell lymphoma, immunocytoma, and small B-cell lymphocytic lymphoma.
J Clin Oncol.
2000;18:317-324
29.
Weng WK, Levy R.
Expression of complement inhibitors CD46, CD55, and CD59 on tumor cells does not predict clinical outcome after rituximab treatment in follicular non-Hodgkin lymphoma.
Blood.
2001;98:1352-1357 30. Frank MM, Fries LF. The role of complement in inflammation and phagocytosis. Immunol Today. 1991;12:322-326[CrossRef][Medline] [Order article via Infotrieve]. 31. Gerard C, Gerard NP. C5A anaphylatoxin and its seven transmembrane-segment receptor. Annu Rev Immunol. 1994;12:775-808[CrossRef][Medline] [Order article via Infotrieve]. 32. Sozzani S, Sallusto F, Luini W, et al. Migration of dendritic cells in response to formyl peptides, C5a, and a distinct set of chemokines. J Immunol. 1995;155:3292-3295[Abstract].
33.
Dobrina A, Pausa M, Fischetti F, et al.
Cytolytically inactive terminal complement complex causes transendothelial migration of polymorphonuclear leukocytes in vitro and in vivo.
Blood.
2002;99:185-192
34.
Colombat P, Salles G, Brousse N, et al.
Rituximab (anti-CD20 monoclonal antibody) as single first-line therapy for patients with follicular lymphoma with a low tumor burden: clinical and molecular evaluation.
Blood.
2001;97:101-106 35. Selenko N, Maidic O, Draxier S, et al. CD20 antibody (C2B8)-induced apoptosis of lymphoma cells promotes phagocytosis by dendritic cells and cross-priming of CD8+ cytotoxic T cells. Leukemia. 2001;15:1619-1626[CrossRef][Medline] [Order article via Infotrieve].
© 2003 by The American Society of Hematology.
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 |
|
 |
|
  D. Dayde, D. Ternant, M. Ohresser, S. Lerondel, S. Pesnel, H. Watier, A. Le Pape, P. Bardos, G. Paintaud, and G. Cartron Tumor burden influences exposure and response to rituximab: pharmacokinetic-pharmacodynamic modeling using a syngeneic bioluminescent murine model expressing human CD20 Blood, April 16, 2009; 113(16): 3765 - 3772. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. P. Hilchey, O. Hyrien, T. R. Mosmann, A. M. Livingstone, J. W. Friedberg, F. Young, R. I. Fisher, R. J. Kelleher Jr, R. B. Bankert, and S. H. Bernstein Rituximab immunotherapy results in the induction of a lymphoma idiotype-specific T-cell response in patients with follicular lymphoma: support for a "vaccinal effect" of rituximab Blood, April 16, 2009; 113(16): 3809 - 3812. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Dalle, S. Dupire, S. Brunet-Manquat, L. Reslan, A. Plesa, and C. Dumontet In vivo Model of Follicular Lymphoma Resistant to Rituximab Clin. Cancer Res., February 1, 2009; 15(3): 851 - 857. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Racila, B. K. Link, W.-K. Weng, T. E. Witzig, S. Ansell, M. J. Maurer, J. Huang, C. Dahle, A. Halwani, R. Levy, et al. A Polymorphism in the Complement Component C1qA Correlates with Prolonged Response Following Rituximab Therapy of Follicular Lymphoma Clin. Cancer Res., October 15, 2008; 14(20): 6697 - 6703. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Fu, D. D. Weisenburger, W. W.L. Choi, K. D. Perry, L. M. Smith, X. Shi, C. P. Hans, T. C. Greiner, P. J. Bierman, R. G. Bociek, et al. Addition of Rituximab to Standard Chemotherapy Improves the Survival of Both the Germinal Center B-Cell-Like and Non-Germinal Center B-Cell-Like Subtypes of Diffuse Large B-Cell Lymphoma J. Clin. Oncol., October 1, 2008; 26(28): 4587 - 4594. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 X. Zhou, W. Hu, and X. Qin The Role of Complement in the Mechanism of Action of Rituximab for B-Cell Lymphoma: Implications for Therapy Oncologist, September 1, 2008; 13(9): 954 - 966. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. Cartron, M. Ohresser, G. Salles, P. Solal-Celigny, P. Colombat, and H. Watier Neutrophil role in in vivo anti-lymphoma activity of rituximab: FCGR3B-NA1/NA2 polymorphism does not influence response and survival after rituximab treatment Ann. Onc., August 1, 2008; 19(8): 1485 - 1487. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Luqman, S. Klabunde, K. Lin, G. V. Georgakis, A. Cherukuri, J. Holash, C. Goldbeck, X. Xu, E. E. Kadel III, S. H. Lee, et al. The antileukemia activity of a human anti-CD40 antagonist antibody, HCD122, on human chronic lymphocytic leukemia cells Blood, August 1, 2008; 112(3): 711 - 720. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. Macor, C. Tripodo, S. Zorzet, E. Piovan, F. Bossi, R. Marzari, A. Amadori, and F. Tedesco In vivo Targeting of Human Neutralizing Antibodies against CD55 and CD59 to Lymphoma Cells Increases the Antitumor Activity of Rituximab Cancer Res., November 1, 2007; 67(21): 10556 - 10563. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
X. Zhao, R. Lapalombella, T. Joshi, C. Cheney, A. Gowda, M. S. Hayden-Ledbetter, P. R. Baum, T. S. Lin, D. Jarjoura, A. Lehman, et al. Targeting CD37-positive lymphoid malignancies with a novel engineered small modular immunopharmaceutical Blood, October 1, 2007; 110(7): 2569 - 2577. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
W. Zhang, M. Gordon, A. M. Schultheis, D. Y. Yang, F. Nagashima, M. Azuma, H.-M. Chang, E. Borucka, G. Lurje, A. E. Sherrod, et al. FCGR2A and FCGR3A Polymorphisms Associated With Clinical Outcome of Epidermal Growth Factor Receptor Expressing Metastatic Colorectal Cancer Patients Treated With Single-Agent Cetuximab J. Clin. Oncol., August 20, 2007; 25(24): 3712 - 3718. [Abstract] [Full Text] [PDF]
|
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 |
|
 P. Dreger, M. Rieger, B. Seyfarth, M. Hensel, M. Kneba, A. D. Ho, N. Schmitz, and C. Pott Rituximab-augmented myeloablation for first-line autologous stem cell transplantation for mantle cell lymphoma: effects on molecular response and clinical outcome Haematologica, January 1, 2007; 92(1): 42 - 49. [Abstract] [Full Text] [PDF]
|
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D. H. Kim, H. D. Jung, J. G. Kim, J.-J. Lee, D.-H. Yang, Y. H. Park, Y. R. Do, H. J. Shin, M. K. Kim, M. S. Hyun, et al. FCGR3A gene polymorphisms may correlate with response to frontline R-CHOP therapy for diffuse large B-cell lymphoma Blood, October 15, 2006; 108(8): 2720 - 2725. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
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 R. M. Sharkey and D. M. Goldenberg Targeted Therapy of Cancer: New Prospects for Antibodies and Immunoconjugates CA Cancer J Clin, July 1, 2006; 56(4): 226 - 243. [Abstract] [Full Text] [PDF]
|
|
|
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T. van Meerten, R. S. van Rijn, S. Hol, A. Hagenbeek, and S. B. Ebeling Complement-Induced Cell Death by Rituximab Depends on CD20 Expression Level and Acts Complementary to Antibody-Dependent Cellular Cytotoxicity. Clin. Cancer Res., July 1, 2006; 12(13): 4027 - 4035. [Abstract] [Full Text] [PDF]
|
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 M. Alfonso-Perez, S. Lopez-Giral, N. E. Quintana, J. Loscertales, P. Martin-Jimenez, and C. Munoz Anti-CCR7 monoclonal antibodies as a novel tool for the treatment of chronic lymphocyte leukemia J. Leukoc. Biol., June 1, 2006; 79(6): 1157 - 1165. [Abstract] [Full Text] [PDF]
|
|
|
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S. Iida, H. Misaka, M. Inoue, M. Shibata, R. Nakano, N. Yamane-Ohnuki, M. Wakitani, K. Yano, K. Shitara, and M. Satoh Nonfucosylated Therapeutic IgG1 Antibody Can Evade the Inhibitory Effect of Serum Immunoglobulin G on Antibody-Dependent Cellular Cytotoxicity through its High Binding to Fc{gamma}RIIIa. Clin. Cancer Res., May 1, 2006; 12(9): 2879 - 2887. [Abstract] [Full Text] [PDF]
|
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|
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C. Carlo-Stella, M. Di Nicola, M. C. Turco, L. Cleris, C. Lavazza, P. Longoni, M. Milanesi, M. Magni, M. Ammirante, A. Leone, et al. The Anti-Human Leukocyte Antigen-DR Monoclonal Antibody 1D09C3 Activates the Mitochondrial Cell Death Pathway and Exerts a Potent Antitumor Activity in Lymphoma-Bearing Nonobese Diabetic/Severe Combined Immunodeficient Mice Cancer Res., February 1, 2006; 66(3): 1799 - 1808. [Abstract] [Full Text] [PDF]
|
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J. F. DiJoseph, M. M. Dougher, L. B. Kalyandrug, D. C. Armellino, E. R. Boghaert, P. R. Hamann, J. K. Moran, and N. K. Damle Antitumor Efficacy of a Combination of CMC-544 (Inotuzumab Ozogamicin), a CD22-Targeted Cytotoxic Immunoconjugate of Calicheamicin, and Rituximab against Non-Hodgkin's B-Cell Lymphoma Clin. Cancer Res., January 1, 2006; 12(1): 242 - 249. [Abstract] [Full Text] [PDF]
|
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|
 C. M. Jubala, J. W. Wojcieszyn, V. E. O. Valli, D. M. Getzy, S. P. Fosmire, D. Coffey, D. Bellgrau, and J. F. Modiano CD20 Expression in Normal Canine B Cells and in Canine non-Hodgkin Lymphoma Vet. Pathol., July 1, 2005; 42(4): 468 - 476. [Abstract] [Full Text] [PDF]
|
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J. M. Pagel, C. Laugen, L. Bonham, R. C. Hackman, D. M. Hockenbery, R. Bhatt, D. Hollenback, H. Carew, J. W. Singer, and O. W. Press Induction of Apoptosis Using Inhibitors of Lysophosphatidic Acid Acyltransferase-{beta} and Anti-CD20 Monoclonal Antibodies for Treatment of Human Non-Hodgkin's Lymphomas Clin. Cancer Res., July 1, 2005; 11(13): 4857 - 4866. [Abstract] [Full Text] [PDF]
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S. D. Gillies, Y. Lan, S. Williams, F. Carr, S. Forman, A. Raubitschek, and K.-M. Lo An anti-CD20-IL-2 immunocytokine is highly efficacious in a SCID mouse model of established human B lymphoma Blood, May 15, 2005; 105(10): 3972 - 3978. [Abstract] [Full Text] [PDF]
|
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|
 P. Carter and C. F. McDonagh Designer Antibody-Based Therapeutics for Oncology Am. Assoc. Cancer Res. Educ. Book, April 1, 2005; 2005(1): 147 - 154. [Full Text] [PDF]
|
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|
|
|
|
G. Lenz, M. Dreyling, E. Hoster, B. Wormann, U. Duhrsen, B. Metzner, H. Eimermacher, A. Neubauer, H. Wandt, H. Steinhauer, et al. Immunochemotherapy With Rituximab and Cyclophosphamide, Doxorubicin, Vincristine, and Prednisone Significantly Improves Response and Time to Treatment Failure, But Not Long-Term Outcome in Patients With Previously Untreated Mantle Cell Lymphoma: Results of a Prospective Randomized Trial of the German Low Grade Lymphoma Study Group (GLSG) J. Clin. Oncol., March 20, 2005; 23(9): 1984 - 1992. [Abstract] [Full Text] [PDF]
|
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|
 |
|
 S A Chambers and D Isenberg Anti-B cell therapy (Rituximab) in the treatment of autoimmune diseases Lupus, March 1, 2005; 14(3): 210 - 214. [Abstract] [PDF]
|
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|
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 |
|
 H. Nehme-Schuster, A.-S. Korganow, J.-L. Pasquali, and T. Martin Rituximab inefficiency during type I cryoglobulinaemia Rheumatology, March 1, 2005; 44(3): 410 - 411. [Full Text] [PDF]
|
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 F. D. Arditti, A. Rabinkov, T. Miron, Y. Reisner, A. Berrebi, M. Wilchek, and D. Mirelman Apoptotic killing of B-chronic lymphocytic leukemia tumor cells by allicin generated in situ using a rituximab-alliinase conjugate Mol. Cancer Ther., February 1, 2005; 4(2): 325 - 332. [Abstract] [Full Text] [PDF]
|
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M. Z. Lin, M. A. Teitell, and G. J. Schiller The Evolution of Antibodies into Versatile Tumor-Targeting Agents Clin. Cancer Res., January 1, 2005; 11(1): 129 - 138. [Abstract] [Full Text] [PDF]
|
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M. L. Palomba, W. K. Roberts, T. Dao, G. Manukian, J. A. Guevara-Patino, J. D. Wolchok, D. A. Scheinberg, and A. N. Houghton CD8+ T-Cell-Dependent Immunity Following Xenogeneic DNA Immunization against CD20 in a Tumor Challenge Model of B-Cell Lymphoma Clin. Cancer Res., January 1, 2005; 11(1): 370 - 379. [Abstract] [Full Text] [PDF]
|
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J. F. DiJoseph, M. E. Goad, M. M. Dougher, E. R. Boghaert, A. Kunz, P. R. Hamann, and N. K. Damle Potent and Specific Antitumor Efficacy of CMC-544, a CD22-Targeted Immunoconjugate of Calicheamicin, against Systemically Disseminated B-Cell Lymphoma Clin. Cancer Res., December 15, 2004; 10(24): 8620 - 8629. [Abstract] [Full Text] [PDF]
|
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G. Cartron, H. Watier, J. Golay, and P. Solal-Celigny From the bench to the bedside: ways to improve rituximab efficacy Blood, November 1, 2004; 104(9): 2635 - 2642. [Abstract] [Full Text] [PDF]
|
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C. F. Eisenbeis, A. Grainger, B. Fischer, R. A. Baiocchi, L. Carrodeguas, S. Roychowdhury, L. Chen, A. L. Banks, T. Davis, D. Young, et al. Combination Immunotherapy of B-Cell Non-Hodgkin's Lymphoma with Rituximab and Interleukin-2: A Preclinical and Phase I Study Clin. Cancer Res., September 15, 2004; 10(18): 6101 - 6110. [Abstract] [Full Text] [PDF]
|
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R. P. Taylor CD20 Trek: the next generation Blood, September 15, 2004; 104(6): 1592 - 1592. [Full Text] [PDF]
|
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|
|
|
J. L. Teeling, R. R. French, M. S. Cragg, J. van den Brakel, M. Pluyter, H. Huang, C. Chan, P. W. H. I. Parren, C. E. Hack, M. Dechant, et al. Characterization of new human CD20 monoclonal antibodies with potent cytolytic activity against non-Hodgkin lymphomas Blood, September 15, 2004; 104(6): 1793 - 1800. [Abstract] [Full Text] [PDF]
|
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|
 |
|
 A. J. Olszewski and M. L. Grossbard Empowering Targeted Therapy: Lessons from Rituximab Sci. Signal., July 13, 2004; 2004(241): pe30 - pe30. [Abstract] [Full Text] [PDF]
|
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S. Dall'Ozzo, S. Tartas, G. Paintaud, G. Cartron, P. Colombat, P. Bardos, H. Watier, and G. Thibault Rituximab-Dependent Cytotoxicity by Natural Killer Cells: Influence of FCGR3A Polymorphism on the Concentration-Effect Relationship Cancer Res., July 1, 2004; 64(13): 4664 - 4669. [Abstract] [Full Text] [PDF]
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M. S. Cragg and M. J. Glennie Antibody specificity controls in vivo effector mechanisms of anti-CD20 reagents Blood, April 1, 2004; 103(7): 2738 - 2743. [Abstract] [Full Text] [PDF]
|
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J. F. DiJoseph, D. C. Armellino, E. R. Boghaert, K. Khandke, M. M. Dougher, L. Sridharan, A. Kunz, P. R. Hamann, B. Gorovits, C. Udata, et al. Antibody-targeted chemotherapy with CMC-544: a CD22-targeted immunoconjugate of calicheamicin for the treatment of B-lymphoid malignancies Blood, March 1, 2004; 103(5): 1807 - 1814. [Abstract] [Full Text] [PDF]
|
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|
|
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C. F. Eisenbeis, M. A. Caligiuri, and J. C. Byrd Rituximab: Converging Mechanisms of Action in Non-Hodgkin's Lymphoma? Clin. Cancer Res., December 1, 2003; 9(16): 5810 - 5812. [Full Text] [PDF]
|
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W.-K. Weng and R. Levy Two Immunoglobulin G Fragment C Receptor Polymorphisms Independently Predict Response to Rituximab in Patients With Follicular Lymphoma J. Clin. Oncol., November 1, 2003; 21(21): 3940 - 3947. [Abstract] [Full Text] [PDF]
|
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|
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H. T. C. Chan, D. Hughes, R. R. French, A. L. Tutt, C. A. Walshe, J. L. Teeling, M. J. Glennie, and M. S. Cragg CD20-induced Lymphoma Cell Death Is Independent of Both Caspases and Its Redistribution into Triton X-100 Insoluble Membrane Rafts Cancer Res., September 1, 2003; 63(17): 5480 - 5489. [Abstract] [Full Text] [PDF]
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 N. Di Gaetano, E. Cittera, R. Nota, A. Vecchi, V. Grieco, E. Scanziani, M. Botto, M. Introna, and J. Golay Complement Activation Determines the Therapeutic Activity of Rituximab In Vivo J. Immunol., August 1, 2003; 171(3): 1581 - 1587. [Abstract] [Full Text] [PDF]
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A. Tzankov, J. Krugmann, F. Fend, M. Fischhofer, R. Greil, and S. Dirnhofer Prognostic Significance of CD20 Expression in Classical Hodgkin Lymphoma: A Clinicopathological Study of 119 Cases Clin. Cancer Res., April 1, 2003; 9(4): 1381 - 1386. [Abstract] [Full Text] [PDF]
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Top
Abstract
Introduction
constant regions, and specific to the
CD20 antigen expressed on mature B lymphocytes.
receptor
(Fc
97%)
of tumor suspensions was evaluated by flow cytometric analysis and
cells were subsequently cryopreserved in liquid nitrogen. They included
7 follicular lymphomas (FLs), 7 mantle cell lymphomas (MCLs), 7 diffuse
large cell lymphomas (DLCLs), 7 small lymphocytic lymphomas
(SLLs), according to the Revised European-American
Classification of Lymphoid Neoplasms (REAL)
classification.
