Ada Schollenberger

Publications of Ada Schollenberger

• Mannosylated lipoarabinomannan balances apoptosis and inflammatory state in mycobacteria-infected and uninfected bystander macrophages.

  Authors: Bartosz Wojtas, Beata Fijalkowska, Adam Wlodarczyk, Ada Schollenberger, Marek Niemialtowski, Beston Hamasur, Andrzej Pawlowski, Malgorzata Krzyzowska

  Microbial pathogenesis. 03/2011; 51(1-2):9-21.

  To assess the role of mannosylated lipoarabinomannan (ManLAM) in the inflammatory and apoptotic response of mycobacteria-infected and uninfected, bystander cells we applied a mouse macrophage modelTo assess the role of mannosylated lipoarabinomannan (ManLAM) in the inflammatory and apoptotic response of mycobacteria-infected and uninfected, bystander cells we applied a mouse macrophage model of infection with avirulent strains--Mycobacterium bovis BCG, Mycobacterium tuberculosis (MTB) H37Ra and compared with a virulent MTB H37Rv strain infection. ManLAM contributed to the infection of macrophages by protection from apoptosis with stabilized Bcl-2 expression and down-regulated Bax expression for infected cells (BCG) or with stabilized Bcl-2 expression for uninfected bystander target cells (H37Ra). Additionally, ManLAM up-regulated FasL expression on the infected cells. Active extracellular signal-regulated kinase (ERK1/2) in BCG and H37Rv infection provided an anti-apoptotic effect by stabilization of anti-apoptotic Bcl-2 expression in the infected cells. Inhibitors specific for c-Jun-NH2-terminal kinase or stress-activated kinase (JNK) and p38 kinase decreased apoptosis of infected cells (BCG, H37Ra) and of uninfected bystanders (H37Ra) by down-regulating Bax. ManLAM significantly down-regulated production of pro-inflammatory IL-12 and TNF-alpha and activation of JNK by both avirulent strains. We conclude that by stabilization of Bcl-2 expression, down-regulation of JNK activity and down-regulation of pro-inflammatory cytokines production ManLAM can contribute to suppression of apoptosis and inflammatory reaction of uninfected, bystander cells.

• [Molecular recognition of viral infections - immune response stimulation].

  Authors: Małgorzata Gieryńska, Ada Schollenberger

  Postȩpy higieny i medycyny doświadczalnej (Online). 01/2011; 65:299-313.

  The mammalian immune system has evolved several mechanisms that allow bacterial and viral infections to be successfully fought. Animal cells are able to recognize viral infection and this recognitionThe mammalian immune system has evolved several mechanisms that allow bacterial and viral infections to be successfully fought. Animal cells are able to recognize viral infection and this recognition is dependent on the presence of intracellular sensors that instantly identify danger signals and initiate signal cascades leading to an effective antiviral response. Several host proteins have been identified as intracellular sensors, namely: Toll-like receptors, RIG-I-like receptors, AIM2-like receptors and DAI, DNA-dependent activator of IFN regulatory factor. They recognize and bind viral genomic nucleic acids and all their replicative intermediates. Receptor-ligand interaction leads to activation of specific metabolic pathways that include synthesis and release of type I interferons and proinflammatory cytokines. These mediators are in turn responsible for synchronizing mechanisms of innate and adaptive antiviral immunity. They are crucial for blocking viral replication, preventing the spread of infection and eventually eliminating the virus from the host. Signaling pathways dependent on RIG-I, independent of TLR and other viral ligand(s) identification mechanisms leading to antiviral immune response stimulation, are discussed in this review.

• [Dendritic epidermal T cells: sentinels of the skin]

  Authors: Małgorzata Gieryńska, Ada Schollenberger, Irma Spohr Cespedes, Ewelina Pawlak

  Postȩpy higieny i medycyny doświadczalnej (Online). 02/2009; 63:114-22.

  Dendritic epidermal T cells (DETCs) are T lymphocytes that express a canonical gammadeltaTCR and form a dense network in murine skin. The major difference between ab and gammadeltaT cells is that theDendritic epidermal T cells (DETCs) are T lymphocytes that express a canonical gammadeltaTCR and form a dense network in murine skin. The major difference between ab and gammadeltaT cells is that the latter do not require antigen presentation in the context of MHC I or II for stimulation. Using their gammadeltaTCR they recognize so far unknown ligands expressed by stressed, infected, or transformed keratinocytes. Since DETCs are located only in the skin, they provide the front line of defense against invasion, but also take part in immune regulation. These cells are in intimate contact with neighboring cells and, through their unique antigen recognition, can immediately react to incoming signals and secrete a variety of cytokines, growth factors, and chemokines that have been implicated in tissue repair and cell survival, proliferation, migration, and recruitment. They play a large role in maintaining skin homeostasis by rapid induction of immune response and at the same time they can release immunoregulatory mediators that inhibit inflammation and consequently maintain integrity of the skin.

• Dendritic epidermal T cells: their role in the early phase of ectromelia virus infection.

  Authors: Małgorzata Gieryńska, Ewelina Pawlak, Ada Schollenberger, Irma Spohr Cespedes

  Postȩpy higieny i medycyny doświadczalnej (Online). 02/2009; 63:369-76.

  INTRODUCTION: Dendritic epidermal T cells (DETCs) are bone marrow-derived T lymphocytes that express a canonical gammadelta TCR and form a dense network in the murine skin. Here, we sought toINTRODUCTION: Dendritic epidermal T cells (DETCs) are bone marrow-derived T lymphocytes that express a canonical gammadelta TCR and form a dense network in the murine skin. Here, we sought to determine their role during the early phase of ectromelia virus (ECTV) infection. MATERIALS/METHODS: In vivo and in vitro models were established for this purpose. In the first model, C57BL/6 mice were intradermally infected into the central part of the ear pinnas with 105 PFU ECTV-Mos strain per ear. At indicated time-points, the total pinna cell population was isolated to determine the presence of DETCs and the enumeration of DETCs secreting IFN-gamma under in vitro stimulation. Purified DETCs were also analyzed for certain gene expressions by RT-PCR. In the second model, purified DETCs isolated from pinnas of uninfected C57BL/6 mice were stimulated in vitro with 5 MOI of UV-inactivated ECTV-Mos. Total RNA was isolated at indicated time-points for RT-PCR gene expression evaluation. RESULTS: A rapid increase in DETCs number in the pinnas was observed for 24 hours post-infection. During the next 24 h the DETCs number decreased, reaching control values. Rapid but short-lasting INF-gamma secretion by purified DETCs in vitro was observed and correlated well with the expression of the beta chemokine CCL5 gene responsible for macrophage and neutrophil attraction. It was also accompanied by DETCs expression of the immunoregulatory factors TGF-beta, GM-CSF, and KGF genes important for maintaining skin integrity. CONCLUSIONS: DETCs from mice infected with ECTV-Mos were rapidly induced to cascade the secretion of mediators that contribute to both immune protection and the control of skin integrity.

• Gene expression profiling of lipoarabinomannan-treated mouse macrophage cultures infected with Mycobacterium bovis BCG.

  Authors: Małgorzata Krzyzowska, Tadeusz Malewski, Beston Hamasur, Ewa Augustynowicz-Kopeć, Ada Schollenberger, Andrzej Pawłowski, Marek Niemiałtowski

  Polish journal of microbiology / Polskie Towarzystwo Mikrobiologów = The Polish Society of Microbiologists. 01/2008; 57(2):125-33.

  The mannosylated lipoarabinomanan (ManLAM) from mycobacterial species possesses strong anti-apoptotic action. Here we examined the ability of ManLAM isolated from Mycobacterium tuberculosis H37Rv toThe mannosylated lipoarabinomanan (ManLAM) from mycobacterial species possesses strong anti-apoptotic action. Here we examined the ability of ManLAM isolated from Mycobacterium tuberculosis H37Rv to alter expression profiles of apoptosis-related genes in mouse macrophages infected with Mycobacterium bovis BCG Danish strain. ManLAM suppressed BCG-induced apoptosis and activities of caspase-1, -3, -8 and 9. Mouse Apoptosis Gene Array showed that ManLAM significantly down-regulated pro-apoptotic and proinflammatory genes: caspase-1, -3, -7, -8 and -9, TNF-alpha/TNFSF2, Fas/TNFRSF6, Bax-alpha, as well as IL-12 p35 and iNOS simultaneously up-regulating anti-apoptotic genes such as Bcl-2 and Mcl-1. The effect of ManLAM was contrary to BCG-induced up-regulation of proapoptotic and pro-inflammatory genes and consistent with the functional data.

• Lipoarabinomannan as a regulator of the monocyte apoptotic response to Mycobacterium bovis BCG Danish strain 1331 infection.

  Authors: Małgorzata Krzyzowska, Ada Schollenberger, Andrzej Pawłowski, Beston Hamasur, Anna Winnicka, Ewa Augustynowicz-Kopeć, Marek Niemiałtowski

  Polish journal of microbiology / Polskie Towarzystwo Mikrobiologów = The Polish Society of Microbiologists. 02/2007; 56(2):89-96.

  The mannosylated lipoarabinomannan (ManLAM) from mycobacterial species possesses strong immunomodulatory effects. Here we examined the ability of Mycobacterium tuberculosis ManLAM to interfere withThe mannosylated lipoarabinomannan (ManLAM) from mycobacterial species possesses strong immunomodulatory effects. Here we examined the ability of Mycobacterium tuberculosis ManLAM to interfere with the apoptotic response of mouse monocyte cell line, RAW 264.7 infected with Mycobacterium bovis BCG Danish strain. Incubation of BCG-infected monocytes with ManLAM decreased production of NO and the numbers of apoptotic cells which synergized with the polarization of mitochondrial membrane. Activities of caspase-1, -3, -8 and 9 followed pattern of apoptosis suppression by ManLAM, except for caspase-1, which showed no significant change in activity. ManLAM also stabilized anti-apoptotic ratio of bcl-2/bax expression in BCG-infected cells and blocked activation of Fas/FasL-induced pathway of apoptosis. Thus, ManLAM, apart from blocking mitochondrial pathway of apoptosis, may induce several other pathways regulating apoptotic response in BCG-infected mouse monocytes.

• Mousepox conjunctivitis: the role of Fas/FasL-mediated apoptosis of epithelial cells in virus dissemination.

  Authors: Malgorzata Krzyzowska, Magdalena Polanczyk, Monika Bas, Joanna Cymerys, Ada Schollenberger, Francesca Chiodi, Marek Niemialtowski

  The Journal of general virology. 08/2005; 86(Pt 7):2007-18.

  BALB/c mice infected with the Moscow strain of Ectromelia virus (ECTV-MOS) show a large number of apoptotic cells, and an influx of lymphoid cells in the epithelium and substantia propria ofBALB/c mice infected with the Moscow strain of Ectromelia virus (ECTV-MOS) show a large number of apoptotic cells, and an influx of lymphoid cells in the epithelium and substantia propria of conjunctivae, respectively. The presence of ECTV-MOS antigens in the epithelium of conjunctivae significantly upregulates Fas in the epithelial layer and FasL in the suprabasal layer of conjunctiva. Inhibition of FasL with blocking antibodies in cultures of conjunctival cells isolated from ECTV-MOS-infected BALB/c mice showed that the Fas/FasL pathway is important in apoptosis of ECTV-MOS-infected cells. The results also showed that the presence of cytokines, in particular interferon (IFN)-gamma, upregulated expression of Fas. Interleukin (IL) 2, 4, 10 and IFN-gamma were produced at the peak of conjunctivitis (at day 15 of infection) with a predominance of IFN-gamma and a small, but significant, production of IL4 and IL10 compared with non-infected animals. These results suggest that not only is Fas/FasL expression in conjunctiva involved in elimination of migrating Fas+ cells but also plays an important role in the turnover of conjunctival epithelium and thus may be crucial for ECTV spreading to the surrounding environment.

• Apoptosis during ectromelia orthopoxvirus infection is DEVDase dependent: in vitro and in vivo studies.

  Authors: Malgorzata Krzyzowska, Ada Schollenberger, Janusz Skierski, Marek Niemialtowski

  Microbes and infection / Institut Pasteur. 06/2002; 4(6):599-611.

  Ectromelia virus (EV), which causes mousepox, is a member of the orthopoxviruses that are defined as being able to suppress apoptosis. Caspase-3 is one of the key effector proteases which regulatesEctromelia virus (EV), which causes mousepox, is a member of the orthopoxviruses that are defined as being able to suppress apoptosis. Caspase-3 is one of the key effector proteases which regulates the apoptotic cascade and which is responsible for DNA fragmentation observed during apoptosis. It is well known that viruses, especially poxviruses, can inhibit caspase activity. Here, we report that EV can regulate apoptosis in vitro, suppressing the activity of caspases recognizing the DEVD (Asp-Glu-Val-Asp) motif (caspase-3 and -7) before successful virus replication is completed. Caspase-3 activity measurement showed that an increase in caspase-3 activity preceded the peak of DNA fragmentation demonstrated by TUNEL staining of L929 and RK-13 cells. By using specific caspase inhibitors (Ac-DEVD-CHO, Ac-IETD-CHO and zVAD-fmk), we showed that caspase-3 and -7 (DEVDases) are major effector caspases during EV-induced apoptosis in permissive L929 and RK-13 cell cultures. Apoptosis in vivo seems to play an important role during viraemia as well as during the clearance of EV from genetically susceptible BALB/c (H-2(d)) mice. However, as shown by measurement of caspase-3 activity, caspase-3 protein detection and M30-antibody staining, both DEVDases seem to play an important role during EV clearance from draining lymph nodes and conjunctivae at 15 days p.i. up to 20 days p.i., whereas in the liver and spleen DNA fragmentation coexisted with viral multiplication and secondary viraemia. Apoptosis was DEVDase dependent only in the liver, while spleen DNA fragmentation observed between 5 and 10 days p.i. was caspase independent. Therefore, we conclude that DEVDase- (caspase-3- and caspase-7-) dependent apoptosis is an important mechanism regulating the resolution of EV infection.

• Apoptosis during ectromelia orthopoxvirus infection is DEVDase dependent: in vitro and in vivo studies

  Authors: Małgorzata Krzyżowska, Ada Schollenberger, Janusz Skierski, Marek Niemiałtowski

  Microbes and Infection.

  Ectromelia virus (EV), which causes mousepox, is a member of the orthopoxviruses that are defined as being able to suppress apoptosis. Caspase-3 is one of the key effector proteases which regulatesEctromelia virus (EV), which causes mousepox, is a member of the orthopoxviruses that are defined as being able to suppress apoptosis. Caspase-3 is one of the key effector proteases which regulates the apoptotic cascade and which is responsible for DNA fragmentation observed during apoptosis. It is well known that viruses, especially poxviruses, can inhibit caspase activity. Here, we report that EV can regulate apoptosis in vitro, suppressing the activity of caspases recognizing the DEVD (Asp–Glu–Val–Asp) motif (caspase-3 and -7) before successful virus replication is completed. Caspase-3 activity measurement showed that an increase in caspase-3 activity preceded the peak of DNA fragmentation demonstrated by TUNEL staining of L929 and RK-13 cells. By using specific caspase inhibitors (Ac-DEVD–CHO, Ac-IETD–CHO and zVAD–fmk), we showed that caspase-3 and -7 (DEVDases) are major effector caspases during EV-induced apoptosis in permissive L929 and RK-13 cell cultures. Apoptosis in vivo seems to play an important role during viraemia as well as during the clearance of EV from genetically susceptible BALB/c (H-2d) mice. However, as shown by measurement of caspase-3 activity, caspase-3 protein detection and M30-antibody staining, both DEVDases seem to play an important role during EV clearance from draining lymph nodes and conjunctivae at 15 days p.i. up to 20 days p.i., whereas in the liver and spleen DNA fragmentation coexisted with viral multiplication and secondary viraemia. Apoptosis was DEVDase dependent only in the liver, while spleen DNA fragmentation observed between 5 and 10 days p.i. was caspase independent. Therefore, we conclude that DEVDase- (caspase-3- and caspase-7-) dependent apoptosis is an important mechanism regulating the resolution of EV infection.

• Pathogenesis of mousepox in H-2d mice: evidence for MHC class I-restricted CD8+ and MHC class II-restricted CD4+ CTL antiviral activity in the lymph nodes, spleen and skin, but not in the conjunctivae

  Authors: Irma Spohr Cespedes, Felix N. Toka, Ada Schollenberger, Małgorzata Gieryńska, Marek Niemiałtowski

  Microbes and Infection.

  Genetically sensitive mice (i.e. H-2d haplotype) infected with a natural mouse pathogen named ectromelia virus (EV) can develop a mousepox. Virus replicates well in the skin, next in the drainingGenetically sensitive mice (i.e. H-2d haplotype) infected with a natural mouse pathogen named ectromelia virus (EV) can develop a mousepox. Virus replicates well in the skin, next in the draining lymph nodes (DLNs) and then in the spleen and liver, where it may induce extensive necrosis with strong inflammatory reaction. It is well known from the studies defined on some other viruses that a correlation, functional link and powerful help exist between MHC class I-restricted CD8+ and MHC class II-restricted CD4+ virus-specific cytotoxic T lymphocytes (CTLs). However, in the case of mousepox the role of CD4+ CTLs is still controversial and some reports support the notion that induction of EV-specific CD4+ CTLs is nonessential for the generation of virus-specific immune response. Consequently, this study was designed to evaluate EV-specific CD8+ and CD4+ CTL activity in the DLNs, spleen, skin and conjunctivae of BALB/c (H-2d) mice at 7 and 14 days p.i. with Moscow strain of EV. By using bulk cytotoxicity assay and immunosurgery of effector T cells with mAb specific for CD4+ and/or CD8+ T cells our data show that EV-specific CD8+ CTLs predominated in DLNs and spleen at 7 days (67 and 66% of total CTLs, respectively) and 14 days p.i. (63 and 69% of total CTLs, respectively). In contrast, we found that EV clearance from the cutaneous lesions during mousepox is CD4+ CTL-dependent at 7 days p.i. (59% of total CTLs), whereas at 14 days p.i. CD8+ CTLs predominated in the epidermis, accounting for 72% of the total EV-specific CTLs. Our studies showed that the population of EV-specific CTLs is heterogenous and contains cells of both phenotypes: CD8+ and CD4+. However, these effector cells did not express a similar tendency in cytotoxic activity in the DLNs, spleen and skin in comparison to the conjunctivae where EV-specific CD8+ and CD4+ CTLs were not detected at 7 days p.i. and at peak of mousepox conjunctivitis (14 days p.i.). Our results are discussed in terms of the value of EV to study antiviral CTL responses in the genetically susceptible host.