Antigens Protected Functional Red Blood Cells By The Membrane Grafting Of Compact Hyperbranched Polyglycerols

Red blood cell (RBC) transfusion is vital for the treatment of a number of acute and chronic medical problems such as thalassemia major and sickle cell anemia ^1-3. Due to the presence of multitude of antigens on the RBC surface (~308 known antigens ⁴), patients in the chronic blood transfusion therapy develop alloantibodies due to the miss match of minor antigens on transfused RBCs ^{4, 5}. Grafting of hydrophilic polymers such as polyethylene glycol (PEG) and hyperbranched polyglycerol (HPG) forms an exclusion layer on RBC membrane that prevents the interaction of antibodies with surface antigens without affecting the passage of small molecules such as oxygen ,glucose, and ions³. At present no method is available for the generation of universal red blood donor cells in part because of the daunting challenge presented by the presence of large number of antigens (protein and carbohydrate based) on the RBC surface and the development of such methods will significantly improve transfusion safety, and dramatically improve the availability and use of RBCs. In this report, the experiments that are used to develop antigen protected functional RBCs by the membrane grafting of HPG and their characterization are presented. HPGs are highly biocompatible compact polymers ^{6, 7}, and are expected to be located within the cell glycocalyx that surrounds the lipid membrane ^{8, 9} and mask RBC surface antigens^{10, 11}.     

Generation of human-melanoma-specific T lymphocyte clones defining novel cytolytic targets with panels of newly established melanoma cell lines

Melanoma is a cancer where the immune system is believed to play an important role in the control of malignant cell growth. To study the variability of the immune response in melanoma patients, we derived melanoma cell lines from several HLA-A2⁺ and HLA-A2^– patients. The melanoma cell lines studied were designated FM3, FM6, FM9, FM28, FM37, FM45, FM55_P, FM55_M1 and FM55_M2 and were established from eight metastatic tumors as well as from one primary tumor from a total of seven different patients. On the basis of the ability of tumor cells to induce specific cytotoxic T lymphocytes (CTL) from peripheral blood lymphocytes (PBL) in mixed lymphocyte/tumor culture with HLA-A2⁺ melanoma cells, the FM3 cell line was characterized as highly immunogenic. To investigate the expression of different melanoma-associated antigens recognized by CTL on different melanoma cell lines, we selected the cell line FM3 for restimulation and further T cell cloning experiments. The lytic activity of CTL clones with good proliferative activity was examined using a panel of HLA-A2⁺ and HLA-A2^– melanoma cell lines. None of the tested HLA-A2^– melanoma cell lines were susceptible to lysis by the CTL clones, whereas allogeneic HLA-A2⁺ melanoma cell lines were lysed only by a few CTL clones. On the basis of their reactivity with different melanoma cell lines, it was possible to divide the present CTL clones into at least four groups suggesting the recognition of at least four different antigens. Three of these target structures probably are different from already-described HLA-A2-restricted melanoma-associated antigens, because their expression in the different melanoma cell lines do not correlate with the recognition of melanoma cells by these CTL. The results first indicate that poorly immunogenic melanoma cells may express melanoma-associated antigens, and also suggest that, by using CTL clones obtained against different HLA-class-I-matched melanoma cells, it is possible to define such antigens.     

Lymphoid Cell Lines: Activation of a Latent Herpes-like Virus Particle?

SINCE the discovery¹ of the herpes-like virus particle (EBV) in cell cultures of Burkitt's lymphoma, the virus has been found in many (but not all) lymphoid cell cultures derived from patients with Burkitt's lymphoma² and other lymphoid neoplasms³ or infectious mononucleosis⁴ as well as from normal individuals⁵. The virus has also been found in some biopsies of Burkitt tumours⁶. Serological evidence has implicated it as a causal or associated agent in heterophile-positive infectious mononucleosis^{7, 8}, although its relationship to Burkitt's lymphoma is less clear. Antigenic studies suggest a relationship between the herpes-like virus and antigens detected by indirect immunofluorescence (IF) techniques in fixed cells⁹, surface antigens on viable cells¹⁰ and complement-fixing (CF) antigens in extracts of Burkitt cell cultures¹¹.     

H-2 antigens incorporated into phospholipid vesicles interact specifically with allogeneic cytotoxic T lymphocytes

Subcellular fractions containing different H-2 antigens were tested for their ability to inhibit specific T cell-target cell conjugate formation. H-2-containing membrane vesicles, lentil-lectin-purified H-2 antigens solubilized with detergent (referred to in the text as high-density fraction) or incorporated into lipid vesicles, inhibited T cell-target cell conjugate formation effectively and specifically. However, two- to threefold more protein was required to inhibit T cell-target cell conjugate formation when detergent-solubilized lentil-lectin-purified H-2 antigens were tested. This suggests that a lipid matrix is advantageous for interaction with anti-H-2 T-cell receptors. Experiments were also undertaken to demonstrate specific binding of liposomes containing ¹²⁵I-labeled H-2 antigen to anti-H-2-specific cytotoxic T lymphocytes (CTLs). The binding of the ¹²⁵I-labeled H-2-containing liposomes was saturable and was specifically inhibited by unlabeled H-2 antigens. Monospecific anti-H-2 sera specifically inhibited the binding of liposomes containing H-2 antigen to the CTLs. The results suggest that a specific interaction can occur between serologically defined H-2 antigens and the receptor of anti-H-2 CTLs.     

Immunological characterization of embryonic cell surface antigens recognized by antiblastocyst serum

Stage-specific cell surface antigens expressed during mouse preimplantation development and detected by a rabbit antiserum prepared against mouse blastocysts (A-BL₂) have been characterized by serological and biochemical techniques. Immunofluorescence, immunoradiolabeling, and complement-mediated cytotoxicity assays reveal the expression of A-BL₂ surface antigens beginning at the 4-cell stage and reaching a maximum at the 8-cell to morula stages. At earlier times in development A-BL₂ antigens are not detectable, and there is a decline in expression at the blastocyst stage. No antibody reactivity is detected against adult mouse tissues or teratocarcinoma cell lines. The presence of A-BL₂ antibodies during in vitro embryo culture interferes with normal development. Treatment of embryos with β-N-acetylglucosaminidase, but not other glycosidases, proteases, or lipases, results in a quantitative decrease in the binding of A-BL₂ antibodies to surface antigens. Immunoprecipitation and electrophoretic analyses of A-BL₂ antigens demonstrate specific antibody activity against a pair of embryonic glycoproteins of 65,000 to 70,000 daltons which can be metabolically labeled with ³⁵S-methionine and ³H-glucosamine. Tunicamycin treatment alters the form of the A-BL₂ immunoprecipitate to a single 60,000-dalton protein.     

Distinct Effects on Diversifying Selection by Two Mechanisms of Immunity against Streptococcus pneumoniae

Antigenic variation to evade host immunity has long been assumed to be a driving force of diversifying selection in pathogens. Colonization by Streptococcus pneumoniae, which is central to the organism''s transmission and therefore evolution, is limited by two arms of the immune system: antibody- and T cell- mediated immunity. In particular, the effector activity of CD4⁺ T_H17 cell mediated immunity has been shown to act in trans, clearing co-colonizing pneumococci that do not bear the relevant antigen. It is thus unclear whether T_H17 cell immunity allows benefit of antigenic variation and contributes to diversifying selection. Here we show that antigen-specific CD4⁺ T_H17 cell immunity almost equally reduces colonization by both an antigen-positive strain and a co-colonized, antigen-negative strain in a mouse model of pneumococcal carriage, thus potentially minimizing the advantage of escape from this type of immunity. Using a proteomic screening approach, we identified a list of candidate human CD4⁺ T_H17 cell antigens. Using this list and a previously published list of pneumococcal Antibody antigens, we bioinformatically assessed the signals of diversifying selection among the identified antigens compared to non-antigens. We found that Antibody antigen genes were significantly more likely to be under diversifying selection than the T_H17 cell antigen genes, which were indistinguishable from non-antigens. Within the Antibody antigens, epitopes recognized by human antibodies showed stronger evidence of diversifying selection. Taken together, the data suggest that T_H17 cell-mediated immunity, one form of T cell immunity that is important to limit carriage of antigen-positive pneumococcus, favors little diversifying selection in the targeted antigen. The results could provide new insight into pneumococcal vaccine design.     

Murine leukemia cell hybrids: The quantity of TL antigens expressed by parental and hybrid cells fails to correlate with their sensitivity to TL antibody and complement

The quantity of thymus-leukemia (TL) antigens expressed by murine leukemia cells is significantly greater than that expressed by somatic hybrids of such cells. Based upon the results of ¹²⁵I-lactoperoxidase labeling and antibody absorption procedures, and corrected for size differences between the two cell types, the quantity of TL antigens expressed by RADA-1 cells, a radiation-induced murine leukemia cell line of strain A/J mice, is approximately 5.0 times greater than that of somatic hybrids of RADA-1 and LM(TK)^? cells. LM(TK)^? cells are a thymidine kinase-deficient TL(-) mouse fibroblast cell line. The quantity of TL antigens expressed is related only in part to their susceptibility to lysis by TL antibodies and guinea pig complement (GPC). RADA-1 cells resist lysis. The quantity of TL antigens expressed by RADA-1 cells is analogous to that formed by nonneoplastic thymocytes obtained from F₁ hybrids of two strains of TL(+) and TL(-) mice; cells from both strains are sensitive to TL antiserum and GPC. ASL-1 cells, a spontaneously occurring leukemia cell line of A/J mice, express TL antigens in significantly higher quantities than any of the cell types examined. Exposed to TL antisera, the quantity of TL antigens of ASL-1 cells, but not that of hybrid cells, gradually diminishes. ASL-1 cells convert over a 6-h period of exposure to antibody and guinea pig complement (GPC) resistance; hybrid cells remain sensitive. However, ASL-1 cells converted to TL antibody and GPC resistance continue for a time to express TL antigens in quantities similar to that of sensitive F₁ thymocytes and resistant RADA-1 cells. RADA-1 X LM(TK)^? hybrid cells, which are sensitive to TL antibodies and GPC, express the lowest quantities of TL antigens of any of the cell types examined. It is likely that differences in the quantities of TL antigens expressed by different cell lines reflect genetic mechanisms controlling TL antigen expression. The failure of TL antisera to affect the quantities of TL antigens expressed by hybrid cells is taken as an indication that genetic controls governing antigen expression may be distinguished from those involved in regulating responsiveness to specific antiserum.     

Human Mycobacterium tuberculosis CD8 T Cell Antigens/Epitopes Identified by a Proteomic Peptide Library

Identification of CD8⁺ T cell antigens/epitopes expressed by human pathogens with large genomes is especially challenging, yet necessary for vaccine development. Immunity to tuberculosis, a leading cause of mortality worldwide, requires CD8⁺ T cell immunity, yet the repertoire of CD8 antigens/epitopes remains undefined. We used integrated computational and proteomic approaches to screen 10% of the Mycobacterium tuberculosis (Mtb) proteome for CD8 Mtb antigens. We designed a weighting schema based upon a Multiple Attribute Decision Making:framework to select 10% of the Mtb proteome with a high probability of containing CD8⁺ T cell epitopes. We created a synthetic peptide library consisting of 15-mers overlapping by 11 aa. Using the interferon-γ ELISPOT assay and Mtb-infected dendritic cells as antigen presenting cells, we screened Mtb-specific CD8⁺ T cell clones restricted by classical MHC class I molecules (MHC class Ia molecules), that were isolated from Mtb-infected humans, against this library. Three novel CD8 antigens were unambiguously identified: the EsxJ family (Rv1038c, Rv1197, Rv3620c, Rv2347c, Rv1792), PE9 (Rv1088), and PE_PGRS42 (Rv2487c). The epitopes are B5701-restricted EsxJ_24–34, B3905-restricted PE9_53–67, and B3514-restricted PE_PGRS42_48–56, respectively. The utility of peptide libraries in identifying unknown epitopes recognized by classically restricted CD8⁺ T cells was confirmed, which can be applied to other intracellular pathogens with large size genomes. In addition, we identified three novel Mtb epitopes/antigens that may be evaluated for inclusion in vaccines and/or diagnostics for tuberculosis.     

Exposure of a Distinct PDCA-1+ (CD317) B Cell Population to Agonistic Anti-4-1BB (CD137) Inhibits T and B Cell Responses Both In Vitro and In Vivo

4-1BB (CD137) is an important T cell activating molecule. Here we report that it also promotes development of a distinct B cell subpopulation co-expressing PDCA-1. 4-1BB is expressed constitutively, and its expression is increased when PDCA-1⁺ B cells are activated. We found that despite a high level of surface expression of 4-1BB on PDCA-1⁺ B cells, treatment of these cells with agonistic anti-4-1BB mAb stimulated the expression of only a few activation markers (B7-2, MHC II, PD-L2), cytokines (IL-12p40/p70), and chemokines (MCP-1, RANTES), as well as sTNFR1, and the immunosuppressive enzyme, IDO. Although the PDCA-1⁺ B cells stimulated by anti-4-1BB expressed MHC II at high levels and took up antigens efficiently, Ig class switching was inhibited when they were pulsed with T-independent (TI) or T-dependent (TD) Ags and adoptively transferred into syngeneic recipients. Furthermore, when anti-4-1BB-treated PDCA-1⁺ B cells were pulsed with OVA peptide and combined with Vα2⁺CD4⁺ T cells, Ag-specific cell division was inhibited both in vitro and in vivo. Our findings suggest that the 4-1BB signal transforms PDCA-1⁺ B cells into propagators of negative immune regulation, and establish an important role for 4-1BB in PDCA-1⁺ B cell development and function.     

Regulation of cytotoxic responses to alloantigens by Ia+ cells

Pretreatment of responder spleen cells with anti-Ia plus complement led to an enhancement of cytotoxic responses to alloantigens as well as to TNP-modified self antigens. This observation confirms previous reports that cytotoxic T lymphocytes (CTL) and their precursors (CLP) are Ia^?. Furthermore, it suggests that the CTL responses to alloantigens or TNP-modified self-antigens are regulated by an Ia⁺ suppressor cell. Absorption studies and studies with anti-Ia sera specific for either the entire I region or the I-E/C subregions suggest that the regulatory cell certainly expresses I-E/C-coded determinants although the possibility that it also expresses I-A/B/J-coded determinats cannot be ruled out. Cell-mixing studies suggest that the regulatory cell is Thy-1^? and requires cell division before it can suppress. A clonal assay for CLP was used to show that the enhancement of the CTL response to alloantigens cannot be accounted for on the basis of an increase in the number of CLP in the anti-Ia + C-treated group.     

Inverse Relationship of Polyoma Tumour Specific Cell Surface Antigen to H-2 Histocompatibility Antigens

NEOPLASTIC transformation is known to be associated with changes in the strength of normal cellular antigens, but the effect can be either an increase or a decrease. In the former category are Forssman antigens in guinea-pig hepatoma¹ and SV40 transformed cells²; HL-A antigens in leukaemic cells³; and “G” antigen in human tumour cells⁴. On the other hand, the intensity of the expression of mouse H-2 histocompatibility antigens is decreased in TL(+) leukaemia⁵ and methylcholanthrene (MCA)-induced tumours⁶. We set out to tell whether the expression of histocompatibility antigens was also affected by transformation with an oncogenic virus and have found that in tumours induced by polyoma virus, the quantity of H-2 antigens varied inversely with the amount of tumour-specific cell surface antigen.     

Requirement for the location of both appropriate and irrelevant H-2 antigens on the same stimulator cell for unspecific DNA-synthesis inhibition by the H-2-antigen-primed,specific suppressor T cells

Specific suppressor T cells (SSTC), primed in vivo with H-2 antigens, have been shown previously to inhibit DNA synthesis in the one-way, three-cell mixed lymphocyte reaction (MLR) provided that (a) the stimulator cells bear the priming H-2 antigens, and (b) the responder cells possessIC+S regions homologous to those of the SSTC. Anti-B10.A BlO.A(2R) SSTC (anti-D^d) and anti-A.AL A.TL SSTC (anti-K^k) are shown here to be able to inhibit the DNA synthesis triggered in MLR, not only by the corresponding antigens, D^d and K^k, respectively, but also by irrelevant, third-party H-2 and Mls products provided that the corresponding and third-party antigens are presented on the same stimulator cell. If stimulatorH-2 regions, whose products interact with SSTC and responders, are located on different stimulator cells within the particular MLR, SSTC activity is not elicited. Participation of cytotoxic T lymphocytes in DNA-synthesis suppression is ruled out. Direct contact or location of the inhibited responder cell very close to SSTC is considered to be required for the development of SSTC activity.     

Modulation of Innate Antigen-Presenting Cell Function by Pre-patent Schistosome Infection

Schistosomes are intravascular helminths that infect over 200 million people worldwide. Deposition of eggs by adult schistosomes stimulates Th2 responses to egg antigens and induces granulomatous pathology that is a hallmark of schistosome infection. Paradoxically, schistosomes require host immune function for their development and reproduction and for egress of parasite eggs from the host. To identify potential mechanisms by which immune cells might influence parasite development prior to the onset of egg production, we assessed immune function in mice infected with developing schistosomes. We found that pre-patent schistosome infection is associated with a loss of T cell responsiveness to other antigens and is due to a diminution in the ability of innate antigen-presenting cells to stimulate T cells. Diminution of stimulatory capacity by schistosome worms specifically affected CD11b⁺ cells and did not require concomitant adaptive responses. We could not find evidence for production of a diffusible inhibitor of T cells by innate cells from infected mice. Rather, inhibition of T cell responsiveness by accessory cells required cell contact and only occurred when cells from infected mice outnumbered competent APCs by more than 3∶1. Finally, we show that loss of T cell stimulatory capacity may in part be due to suppression of IL-12 expression during pre-patent schistosome infection. Modulation of CD4⁺ T cell and APC function may be an aspect of host immune exploitation by schistosomes, as both cell types influence parasite development during pre-patent schistosome infection.     

Failure of Neuraminidase to unmask Histocompatibility Antigens on Trophoblast

DURING outbred pregnancy the mother is exposed to genetically foreign tissue because the offspring inherits transplantation antigens from the father. The survival of the foetus is ensured by the intervention of the trophoblast which does not express transplantation antigens between mother and foetus: mouse trophoblast is not rejected even when transplanted into immune recipients^1,3. The mechanism of this failure to express histocompatibility antigens is not understood^1–4, but Kirby et al. have suggested that the extracellular fibrinoid surrounding trophoblast cells is involved^5,6. Currie has suggested that the thick sialomucinous glycocalyx of the trophoblast cell might “mask” the histocompatibility antigens on the trophoblast^7,8 and has demonstrated that neuraminidase unmasked these antigens⁸. Our experiments, however, show that trophoblast incubated with neuraminidase cannot sensitize allogeneic mice to donor histocompatibility antigens. Furthermore, pretreatment of trophoblastic implants with neuraminidase did not interfere with their proliferation and growth in highly immune allogeneic recipients.     

CD98 Heavy Chain Is a Potent Positive Regulator of CD4+ T Cell Proliferation and Interferon-γ Production In Vivo

Upon their recognition of antigens presented by the MHC, T cell proliferation is vital for clonal expansion and the acquisition of effector functions, which are essential for mounting adaptive immune responses. The CD98 heavy chain (CD98hc, Slc3a2) plays a crucial role in the proliferation of both CD4⁺ and CD8⁺ T cells, although it is unclear if CD98hc directly regulates the T cell effector functions that are not linked with T cell proliferation in vivo. Here, we demonstrate that CD98hc is required for both CD4⁺ T cell proliferation and Th1 functional differentiation. T cell-specific deletion of CD98hc did not affect T cell development in the thymus. CD98hc-deficient CD4⁺ T cells proliferated in vivo more slowly as compared with control T cells. C57BL/6 mice lacking CD98hc in their CD4⁺ T cells could not control Leishmania major infections due to lowered IFN-γ production, even with massive CD4⁺ T cell proliferation. CD98hc-deficient CD4⁺ T cells exhibited lower IFN-γ production compared with wild-type T cells, even when comparing IFN-γ expression in cells that underwent the same number of cell divisions. Therefore, these data indicate that CD98hc is required for CD4⁺ T cell expansion and functional Th1 differentiation in vivo, and suggest that CD98hc might be a good target for treating Th1-mediated immune disorders.     

Lymphocytes express Ia antigens of foreign haplotype following treatment with neuraminidase

Evidence is presented which indicates that neuraminidase (NA) treatment of spleen cells both destroys old Ia antigens and reveals new Ia specificities which are not normally expressed by splenocytes. It was found that NA treatment unmasked alien I-A^k-like specificities on A.TH (I ^s ) spleen cells, and I^s-like antigens on A.TL (I ^k ) spleen cells. These conclusions were based on direct testing of NA-treated targets with a range of alloantisera and on cell-absorption experiments. Furthermore, the cellular distribution of NA-exposed antigens resembled that of convential Ia antigens, the new antigens being expressed on more than 90 percent of splenic B cells and a subpopulation of splenic T cells. However, although some of the antigens exposed by NA on A.TH cells appeared to resemble the Ia. 3 and 15 specificities, additional antigens were involved which did not correlate with any previously described Ia antigens.Sugar inhibition experiments demonstrated the NA-exposed antigens to be carbohydrate in nature, D-galactose being an effective inhibitor in these studies. The proportion of- and-linked D-galactose residues associated with the new antigens depended upon the target cell used and the anti-Ia serum tested. Furthermore, glycolipid extracts from lymphoid cells were shown to contain the NA-exposed antigens.Collectively, these results support the existence of carbohydrate-defined Ia antigens. The simplest interpretation of the findings is that NA clips off terminal sialic acid residues from carbohydrate-defined Ia antigens on the cell surface and exposes subterminal sugars which resemble antigens expressed by otherI-region haplotypes.     

Analysis of hybrids between an H-2+, TL− lymphoma and an H-2+, TL+ lymphoma and its H-2−, TL− variant subline

Hybrids between an H-2⁺, TL⁺ lymphoma and an H-2⁺, TL^– lymphoma were studied for their expression of H-2 and TL antigens. The H-2 antigens of both parents were expressed, the TL specificity of the TL⁺ parent was retained, and the TL specificity characteristic of the mouse strain from which the TL^– lymphoma was derived was not expressed. There was no evidence that the genome of either parent altered the expression of the TL antigens coded for by the genome of the opposite parent. Hybrids between the H-2⁺, TL^– lymphoma and an H-2^–, TL^– variant line derived from the H-2⁺, TL⁺ cell line expressed both theK- andD-regioncoded H-2 antigens and the TL specificity characteristic of the parental cell line from which the variant cell was derived. This result is consistent with the defect in the variant cell's being the result of a mutation affecting a gene coding for a positive element necessary for expression of both TL and the serologically detectable H-2 antigens on the cell surface.     

Cytotoxic T lymphocytes of the rat are predominantly restricted by RTL1.A and Not RT1.C-determined major histocompatibility class I antigens

Two types of biochemically defined class I major histocompatibility complex (MHC) antigens are found in the rat, RTLA antigens that are ubiquitously expressed and RTLC antigens which so far are detectable only on certain cell types, notably B and T lymphocytes. It is shown that the cytotoxic T lymphocyte response to minor H antigens of the LEW strain, including the H-Y antigen, and to TNP-modified syngeneic lymphoid cells is restricted by RTLA but not RTLC gene products. This conclusion is based on bulk culture assays including cold target inhibition tests and limiting dilution experiments using recombinants between the RT1 ^a and RT1 ^u haplotypes. The possibility that class I MHC antigens exist which have no major restriction function is discussed.     

Reduced Number of Transitional and Naive B Cells in Addition to Decreased BAFF Levels in Response to the T Cell Independent Immunogen Pneumovax?23

Protective immunity against T cell independent (TI) antigens such as Streptococcus pneumoniae is characterized by antibody production of B cells induced by the combined activation of T cell independent type 1 and type 2 antigens in the absence of direct T cell help. In mice, the main players in TI immune responses have been well defined as marginal zone (MZ) B cells and B-1 cells. However, the existence of human equivalents to these B cell subsets and the nature of the human B cell compartment involved in the immune reaction remain elusive. We therefore analyzed the effect of a TI antigen on the B cell compartment through immunization of healthy individuals with the pneumococcal polysaccharide (PnPS)-based vaccine Pneumovax^®23, and subsequent characterization of B cell subpopulations. Our data demonstrates a transient decrease of transitional and naïve B cells, with a concomitant increase of IgA⁺ but not IgM⁺ or IgG⁺ memory B cells and a predominant generation of PnPS-specific IgA⁺ producing plasma cells. No alterations could be detected in T cells, or proposed human B-1 and MZ B cell equivalents. Consistent with the idea of a TI immune response, antigen-specific memory responses could not be observed. Finally, BAFF, which is supposed to drive class switching to IgA, was unexpectedly found to be decreased in serum in response to Pneumovax^®23. Our results demonstrate that a characteristic TI response induced by Pneumovax^®23 is associated with distinct phenotypical and functional changes within the B cell compartment. Those modulations occur in the absence of any modulations of T cells and without the development of a specific memory response.