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Treatment with oxoglaucine can enhance host resistance to Candida albicans infection of mice with adjuvant arthritis
Diagnostic Microbiology and Infectious Disease 38 (2000) 17–20
www.elsevier.com/locate/diagmicrobio
Treatment with oxoglaucine can enhance host resistance to Candida albicans infection of mice with adjuvant arthritis Nina Ivanovska, Maria Hristova Department of Immunology, Institute of Microbiology, Bulgarian Academy of Sciences, Sofia, Bulgaria Received 14 March 2000; accepted 13 June 2000
Abstract The alkaloid oxoglaucine reduced CD4⫹ cell clones in adult mice and decreased CD4⫹, CD8⫹ and Ig⫹ levels in newborn mice. It prevented the increase of CD8⫹ and Ig⫹ clones induced by Candida albicans (C. albicans) in adult mice. TNF-␣ serum accumulation was inhibited by oxoglaucine in C. albicans infection and adjuvant arthritis. Treatment with oxoglaucine of arthritic mice, followed by inoculation with C. albicans enhanced the host resistance against the pathogen. © 2000 Elsevier Science Inc. All rights reserved.
1. Introduction The use of antirheumatic drugs in clinical practice is limited because of the undesired side effects they cause. Except the organ damage or dysfunction, their long-term application is attended with impaired host resistance and opportunistic infections are most common complications in immunocompromized patients. Two distinct helper T-cell subsets, known as Th1 and Th2 are claimed to underlay the pathogenesis of several autoimmune disorders as well as host antiinfectious mechanisms. Th1-dominated responses, when remain under control, help to eradicate the infectious agents. Th2-dominated responses are required to suppress the developing autoimmunity. Experimental animal models give the opportunity to investigate how the switching between these two responses changes the course of infectious or immunopathogenic processes. Studies in murine models of C. albicans infection have shown that the increased resistance to C. albicans infection is associated with Th1 cytokines secreted by CD4⫹ cells (Del Sero, 99). The protective Th1 response requires the presence of several cytokines such as IFN- (Cenci et al. 1998), IL-6 (Romani et al. 1996), TNF-␣ (Mencacci et al. 1998), IL-12 (Romani et al. 1994). These cytokines are mainly produced by differentiated CD4⫹ and CD8⫹ subsets, which in many cases work cooperatively (Kerksiek and Pamer, 1999). The factors determining Th1/Th2 polarization appear to be the same for both subsets (Carter and Dutton 1996). The control
E-mail address: [email protected] (N. Ivanovska)
of autoimmune disorders also depends on Th1/Th2 balance (Charlton and Lafferty 1995; Sakaguchi et al. 1996). Does the impaired balance during the arthritic state change the host resistance to infections? Undoubtedly, this theory is helpful to evaluate the interventions based on the modulation of Th1/Th2 balance. In fact, the network existing between Th1- and Th2-mediated cytokines is so complicated, that the exact prediction of the final effect can not yet be achieved. Previously, we have investigated the influence of isoquinoline alkaloid oxoglaucine in mice model of adjuvant-induced arthritis. It was established that the substance affected T- and B-cell immune responses and at a high dose of 10 mg/kg was able to inhibit the developing arthritis (Ivanovska 1997a). The high dose treatment enhanced the susceptibility to Klebsiella pneumoniae and C. albicans infections. At the same time, its application as a second agent after another suppressive drugs, such as cyclophosphamide, indomethacine or prednisolone resulted in restorative effect (Ivanovska 1997b, 2000). The present study is focused on the effect of oxoglaucine on different lymphocyte populations and serum TNF-␣ level in intact and C. albicans infected animals. As well as its influence on the host resistance against C. albicans after administration to arthritic mice was investigated. 2. Materials and methods 2.1. Substance Oxoglaucine was synthesized by oxidation of glaucine with sodium metaperiodate. The substance was dissolved in
0732-8893/00/$ – see front matter © 2000 Elsevier Science Inc. All rights reserved. PII: S 0 7 3 2 - 8 8 9 3 ( 0 0 ) 0 0 1 6 7 - X
18
N. Ivanovska and M. Hristova / Diagnostic Microbiology and Infectious Disease 38 (2000) 17–20
DMSO (final concentration was under 0.01%), further diluted in sterile distilled water and injected i.p. to mice in a volume of 0.2 ml. 2.2. Animals Female mice, inbred strain ICR were housed under standard conditions of temperature (21 ⫾ 2°C), relative humidity (55 ⫾ 10%) and kept on a standard commercial pellet diet with water ad libitum.
Table 1 Phenotype of splenic lymphocytesa Group
CD4⫹
CD8⫹
Noninfected 25.00 ⫾ 0.70f 17.34 ⫾ 0.52 b Candida infection 12.76 ⫾ 0.82** 24.04 ⫾ 0.69** Og 10 mg/kg 10 11.98 ⫾ 0.71** 17.86 ⫾ 0.42 daysc Og 5 mg/kg 3 daysd 8.60 ⫾ 0.48** 14.56 ⫾ 0.93* Og 10 mg/kg ⫺10 to 7.70 ⫾ 0.43** 12.16 ⫾ 0.31** 0 daye
Ig⫹ 26.48 ⫾ 0.90 39.18 ⫾ 1.33** 22.70 ⫾ 1.20 21.96 ⫾ 0.72* 24.50 ⫾ 1.24
a
Splenic cells (1⫻106) were collected at day 10 of infection. Female mice, inbred strain ICR, 8 weeks (w) of age were i.p. inoculated with 1⫻105 Candida cells. c 8 w old mice treated with oxoglaucine. d 5 days old mice were treated with oxoglaucine and infected at the age of 8 w. e 8 w ICR mice were treated with oxoglaucine and then infected. f Values represent mean ⫾ SEM for n⫽5 animals/group; *p ⬍ 0.01, **p ⬍ 0.001 determined by ANOVA. b
2.3. Phenotypic analysis For phenotypic analysis of splenic lymphocyte subpopulations, spleens were aseptically excised, homogenized and washed in 1.0 ml of phosphate-buffered saline (PBS) containing 0.1% bovine serum albumin (BSA) and 0.05 M NaN3. Cells (1⫻106) were resuspended in 0.1 ml of PBSBSA containing anti-CD4 (Gibco, Gaithersburg, USA), anti-CD8 (Gibco) or isotype control (Coulter Corp., Hialeah, USA) conjugated with fluorescein isothiocyanate (FITC) or phycoerythrin (PE) and incubated for 30 min on ice in the dark. Subsequently, cells were washed with cold PBS-BSA, NaN3, fixed in a 1% paraformaldehyde solution and analyzed by fluorescence-activated cell sorter (FACS). The persentage of stained cells in the total cell population was determined. 2.4. Adjuvant-induced arthritis To induce arthritis, 25 l of a suspension (5 mg/ml) of heat killed and dried Mycobacterium tuberculosis (Difco Labs., Detroit, MI, USA) in paraffin oil was injected subcutaneously into the footpad of the right hind paw. The day of adjuvant injection was regarded as day 0. 2.5. Experimental infection Experimental infection was induced by the use of a virulent clinical isolate 562 of Candida albicans (Institute of Infectious and Parasitic Diseases, Sofia, Bulgaria). Prior to inoculation, the yeast was grown on Sabouraud’s agar for 48 h at 37°C. A standard curve was drawn by plotting the extinction, measured at OD620 against the concentration of the yeast, determined by the number of colony forming units in different suspensions. Cells were washed twice in saline and inoculated at a concentration of 1⫻105 cells per mouse intravenously (i.v.). The infection was monitored daily during 14 days when the number of survivors and mean survival (MST) were determined. 2.6. TNF bioassay TNF concentrations were quantified by cytolytic activity directed against the L929 cell line (NBIMCC 98) sensitive
to TNF-␣. A volume of 0.1 ml of cell suspension (3⫻105 cells/ml) in RPMI 1640 medium (Sigma Chemical Co) supplemented with 100 U/ml penicillin—100 g/ml streptomycin (Sigma Chemical Co) and 10% fetal calf serum (Sigma Chemical Co) was dispensed in a flat-bottom 96well plate (Becton Dickinson) and incubated 18 h at 37°C, 5% CO2/95% air. Then supernatants were discarded and 0.1 ml of appropriately diluted sera in supplemented RPMI 1640 medium were added. After 18 h incubation in the presence of 0.4 g/ml antinomycin D (Calbiochem), the cells were fixed with 5% formaldehyde in PBS, and stained with crystal violet (0.5% in ethanol), resolved in 33% glacial acetic and the plate read at 620 nm (Organon Teknika Reader 530). The concentration of TNF was determined based on a standard curve generated with the recombinant mouse TNF-␣ (Genzyme). The sensitivity of the assay was to 2–5 pg/ml of TNF.
3. Results and discussion The application of oxoglaucine to adult mice significantly reduced CD4⫹ cells but did not affect CD8⫹ and Ig⫹ populations (Table 1). When the administration of the alkaloid started at the age of 5 days, decreased levels of CD4⫹, CD8⫹, and Ig⫹ cells were determined. This result is not surprising as the precursor cells might express higher susceptibility to alkaloid’s action than the already differentiated cell clones in mature mice. In Candida infected mice CD4⫹ population was suppressed contrary to the elevated CD8⫹ and Ig⫹ cell levels. Oxoglaucine prevented Candida-induced increase of these populations. Evidently, the alkaloid is able to influence not only the primary immune response but it can affect the late specific response to the yeast. One of the main cytokines needed for the early
N. Ivanovska and M. Hristova / Diagnostic Microbiology and Infectious Disease 38 (2000) 17–20
19
Fig. 1. Effect of oxoglaucine on serum TNF-␣ level of mice with C. albicans infection or adjuvant arthritis. Data are means ⫾SEM from three determinations of individual sera; n ⫽ 5 animals for each point; **p ⬍ 0.01, ***p ⬍ 0.001 by Student’s t-test calculated to the infected group.
protective response against Candida evasion is TNF-␣. The results show that the course of Candida infection is attended with high serum TNF-␣ level with maximum at first day after inoculation. The pretreatment with oxoglaucine in 10 days before the inoculation suppressed to some extent serum TNF-␣ accumulation at first 5 days of infection (Fig. 1). This downregulation of TNF-␣ most probably prevented the pathogen elimination at the initial period of infection. TNF-␣ is also one of the cytokines related to the development of rheumatoid arthritis (Kollias et al. 1999). Different mode of oxoglaucine action was registered in adjuvant arthritis. The alkaloid failed to change the TNF-␣ accumulation until day 14. At the late period it strongly reduced TNF-␣ level, contrary to the second elevation noted in the arthritic group. These different patterns in both processes pointed that various mechanisms might be triggered in each case and it was of interest to determine the consequences of oxoglaucine application in arthritic state in relation to antiinfectious resistance. When arthritic mice were infected at the peak of inflammation (14 day of disease), they were more prone to C. albicans inoculation than the intact mice (Fig. 2). The application of oxoglaucine during 10 days of developing arthritis resulted in protective effect, since the number of survivors was significantly higher as compared to nontreated with oxoglaucine arthritic animals. These results suggest that the ongoing autoimmune process shifts Th1/ Th2 balance to Th2 response. Oxoglaucine possessed suppressive action on host resistance to C. albicans in intact mice but its effect in arthritic animals was beneficial. Thus the alkaloid seems to be perspective for application in au-
toimmune disorders, as its application at the right time might result in less side effects in concern to antiinfectious resistance.
Fig. 2. Effect of oxoglaucine on the outcome of C. albicans infection in mice with adjuvant artritis. Mice (8 w old) were treated i.p. with 10 mg/kg of oxoglaucine from the day of adjuvant injection (day 0) to day 10. Four days later the i.v. inoculation with 1 ⫻ 105 C. albicans cells was done and the infection was monitored daily during 14 days. *p ⬍ 0.001 by 2 test calculated to the group with infection only.
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N. Ivanovska and M. Hristova / Diagnostic Microbiology and Infectious Disease 38 (2000) 17–20
References Carter, L. L., & Dutton, R. W. (1996). Type 1 and Type 2: A fundamental dichotomy for all T-cell subsets. Curr Opinion Immunol 8, 336 –342. Cenci, E., Mencacci, A., Del Sero, G., Fe` d’Ostiani, C., Mosci, P., Kopf, M., & Romani, L. (1998). IFN- is required for IL-12 responsiveness in mice with Candida albicans infection. J Immunol 161, 3543–3550. Charlton, B., & Lafferty, K. J. (1995). The Th1/Th2 balance in autoimmunity. Curr Opinion Immunol 7, 793–798. Ivanovska, N., Philipov, S., & Georgieva, P. (1997a). Immunopharmacological activity of aporphinoid alkaloid oxoglaucine. Pharmacol Res 35, 267–272. Ivanovska, N., & Philipov, S. (1997b). A low dose immunorestorative effect of aporphinoid alkaloid oxoglaucine on experimentally immunosuppressed and infected mice. Meth Find Exp Clin Pharmacol 19, 579 –583. Ivanovska, N., Hristova, M., & Philipov, S. (2000). Immunosuppression and recovery of drug impaired host resistance against Candida albicans infection by oxoglaucine. Pharmacol Res 41, 99 –105. Kerksiek, K. M., & Pamer, E. G. (1999). T cell responses to bacterial infection. Curr Opinion Immunol 11, 400 – 405.
Kollias, G., Douni, E., Kassiotis, G., & Dimitris, K. (1999). On the role of tumor necrosis factor and receptors in models of multiorgan failure, rheumatoid arthritis, multiple sclerosis and inflammatory bowel disease. Immunol Rev 169, 175–194. Mencacci, A., Cenci, E., Bistoni, F., Bacci, A., Del Sero, G., Montagnoli, C., Fe` d’Ostiani, C., & Romani, L. (1998). Specific and nonspecific immunity to Candida albicans: a lesson from genetically modified animals. Res Immunol 149, 352–361. Romani, L., Mencacci, A., Tonnetti, L., Spaccapelo, R., Cenci, E., Puccetti, P., Wolf, S. F., & Bistoni, F. (1994). Interleukin-12 is both required and prognostic in vivo for T helper type 1 differentiation in murine candidiasis. J Immunol 153, 5157–5175. Romani, L., Mencacci, A., Cenci, E., Spaccapelo, R., Toniatti, C., Puccetti, P., Bistoni, F., & Ploi, V. (1996). Impaired neutrophil response and CD4⫹ T helper cell 1 development in interleukin-6-deficient mice infected with Candida albicans. J Exp Med 183, 1–11. Sakaguchi, S., Toda, M., Asano, M., Itoh, M., Morse, S. S., & Sakaguchi, N. (1996). T cell-mediated maintenance of natural self-tolerance: its breakdown as a possible cause of various autoimmune diseases. J Autoimmun 9, 211–220.
www.elsevier.com/locate/diagmicrobio
Treatment with oxoglaucine can enhance host resistance to Candida albicans infection of mice with adjuvant arthritis Nina Ivanovska, Maria Hristova Department of Immunology, Institute of Microbiology, Bulgarian Academy of Sciences, Sofia, Bulgaria Received 14 March 2000; accepted 13 June 2000
Abstract The alkaloid oxoglaucine reduced CD4⫹ cell clones in adult mice and decreased CD4⫹, CD8⫹ and Ig⫹ levels in newborn mice. It prevented the increase of CD8⫹ and Ig⫹ clones induced by Candida albicans (C. albicans) in adult mice. TNF-␣ serum accumulation was inhibited by oxoglaucine in C. albicans infection and adjuvant arthritis. Treatment with oxoglaucine of arthritic mice, followed by inoculation with C. albicans enhanced the host resistance against the pathogen. © 2000 Elsevier Science Inc. All rights reserved.
1. Introduction The use of antirheumatic drugs in clinical practice is limited because of the undesired side effects they cause. Except the organ damage or dysfunction, their long-term application is attended with impaired host resistance and opportunistic infections are most common complications in immunocompromized patients. Two distinct helper T-cell subsets, known as Th1 and Th2 are claimed to underlay the pathogenesis of several autoimmune disorders as well as host antiinfectious mechanisms. Th1-dominated responses, when remain under control, help to eradicate the infectious agents. Th2-dominated responses are required to suppress the developing autoimmunity. Experimental animal models give the opportunity to investigate how the switching between these two responses changes the course of infectious or immunopathogenic processes. Studies in murine models of C. albicans infection have shown that the increased resistance to C. albicans infection is associated with Th1 cytokines secreted by CD4⫹ cells (Del Sero, 99). The protective Th1 response requires the presence of several cytokines such as IFN- (Cenci et al. 1998), IL-6 (Romani et al. 1996), TNF-␣ (Mencacci et al. 1998), IL-12 (Romani et al. 1994). These cytokines are mainly produced by differentiated CD4⫹ and CD8⫹ subsets, which in many cases work cooperatively (Kerksiek and Pamer, 1999). The factors determining Th1/Th2 polarization appear to be the same for both subsets (Carter and Dutton 1996). The control
E-mail address: [email protected] (N. Ivanovska)
of autoimmune disorders also depends on Th1/Th2 balance (Charlton and Lafferty 1995; Sakaguchi et al. 1996). Does the impaired balance during the arthritic state change the host resistance to infections? Undoubtedly, this theory is helpful to evaluate the interventions based on the modulation of Th1/Th2 balance. In fact, the network existing between Th1- and Th2-mediated cytokines is so complicated, that the exact prediction of the final effect can not yet be achieved. Previously, we have investigated the influence of isoquinoline alkaloid oxoglaucine in mice model of adjuvant-induced arthritis. It was established that the substance affected T- and B-cell immune responses and at a high dose of 10 mg/kg was able to inhibit the developing arthritis (Ivanovska 1997a). The high dose treatment enhanced the susceptibility to Klebsiella pneumoniae and C. albicans infections. At the same time, its application as a second agent after another suppressive drugs, such as cyclophosphamide, indomethacine or prednisolone resulted in restorative effect (Ivanovska 1997b, 2000). The present study is focused on the effect of oxoglaucine on different lymphocyte populations and serum TNF-␣ level in intact and C. albicans infected animals. As well as its influence on the host resistance against C. albicans after administration to arthritic mice was investigated. 2. Materials and methods 2.1. Substance Oxoglaucine was synthesized by oxidation of glaucine with sodium metaperiodate. The substance was dissolved in
0732-8893/00/$ – see front matter © 2000 Elsevier Science Inc. All rights reserved. PII: S 0 7 3 2 - 8 8 9 3 ( 0 0 ) 0 0 1 6 7 - X
18
N. Ivanovska and M. Hristova / Diagnostic Microbiology and Infectious Disease 38 (2000) 17–20
DMSO (final concentration was under 0.01%), further diluted in sterile distilled water and injected i.p. to mice in a volume of 0.2 ml. 2.2. Animals Female mice, inbred strain ICR were housed under standard conditions of temperature (21 ⫾ 2°C), relative humidity (55 ⫾ 10%) and kept on a standard commercial pellet diet with water ad libitum.
Table 1 Phenotype of splenic lymphocytesa Group
CD4⫹
CD8⫹
Noninfected 25.00 ⫾ 0.70f 17.34 ⫾ 0.52 b Candida infection 12.76 ⫾ 0.82** 24.04 ⫾ 0.69** Og 10 mg/kg 10 11.98 ⫾ 0.71** 17.86 ⫾ 0.42 daysc Og 5 mg/kg 3 daysd 8.60 ⫾ 0.48** 14.56 ⫾ 0.93* Og 10 mg/kg ⫺10 to 7.70 ⫾ 0.43** 12.16 ⫾ 0.31** 0 daye
Ig⫹ 26.48 ⫾ 0.90 39.18 ⫾ 1.33** 22.70 ⫾ 1.20 21.96 ⫾ 0.72* 24.50 ⫾ 1.24
a
Splenic cells (1⫻106) were collected at day 10 of infection. Female mice, inbred strain ICR, 8 weeks (w) of age were i.p. inoculated with 1⫻105 Candida cells. c 8 w old mice treated with oxoglaucine. d 5 days old mice were treated with oxoglaucine and infected at the age of 8 w. e 8 w ICR mice were treated with oxoglaucine and then infected. f Values represent mean ⫾ SEM for n⫽5 animals/group; *p ⬍ 0.01, **p ⬍ 0.001 determined by ANOVA. b
2.3. Phenotypic analysis For phenotypic analysis of splenic lymphocyte subpopulations, spleens were aseptically excised, homogenized and washed in 1.0 ml of phosphate-buffered saline (PBS) containing 0.1% bovine serum albumin (BSA) and 0.05 M NaN3. Cells (1⫻106) were resuspended in 0.1 ml of PBSBSA containing anti-CD4 (Gibco, Gaithersburg, USA), anti-CD8 (Gibco) or isotype control (Coulter Corp., Hialeah, USA) conjugated with fluorescein isothiocyanate (FITC) or phycoerythrin (PE) and incubated for 30 min on ice in the dark. Subsequently, cells were washed with cold PBS-BSA, NaN3, fixed in a 1% paraformaldehyde solution and analyzed by fluorescence-activated cell sorter (FACS). The persentage of stained cells in the total cell population was determined. 2.4. Adjuvant-induced arthritis To induce arthritis, 25 l of a suspension (5 mg/ml) of heat killed and dried Mycobacterium tuberculosis (Difco Labs., Detroit, MI, USA) in paraffin oil was injected subcutaneously into the footpad of the right hind paw. The day of adjuvant injection was regarded as day 0. 2.5. Experimental infection Experimental infection was induced by the use of a virulent clinical isolate 562 of Candida albicans (Institute of Infectious and Parasitic Diseases, Sofia, Bulgaria). Prior to inoculation, the yeast was grown on Sabouraud’s agar for 48 h at 37°C. A standard curve was drawn by plotting the extinction, measured at OD620 against the concentration of the yeast, determined by the number of colony forming units in different suspensions. Cells were washed twice in saline and inoculated at a concentration of 1⫻105 cells per mouse intravenously (i.v.). The infection was monitored daily during 14 days when the number of survivors and mean survival (MST) were determined. 2.6. TNF bioassay TNF concentrations were quantified by cytolytic activity directed against the L929 cell line (NBIMCC 98) sensitive
to TNF-␣. A volume of 0.1 ml of cell suspension (3⫻105 cells/ml) in RPMI 1640 medium (Sigma Chemical Co) supplemented with 100 U/ml penicillin—100 g/ml streptomycin (Sigma Chemical Co) and 10% fetal calf serum (Sigma Chemical Co) was dispensed in a flat-bottom 96well plate (Becton Dickinson) and incubated 18 h at 37°C, 5% CO2/95% air. Then supernatants were discarded and 0.1 ml of appropriately diluted sera in supplemented RPMI 1640 medium were added. After 18 h incubation in the presence of 0.4 g/ml antinomycin D (Calbiochem), the cells were fixed with 5% formaldehyde in PBS, and stained with crystal violet (0.5% in ethanol), resolved in 33% glacial acetic and the plate read at 620 nm (Organon Teknika Reader 530). The concentration of TNF was determined based on a standard curve generated with the recombinant mouse TNF-␣ (Genzyme). The sensitivity of the assay was to 2–5 pg/ml of TNF.
3. Results and discussion The application of oxoglaucine to adult mice significantly reduced CD4⫹ cells but did not affect CD8⫹ and Ig⫹ populations (Table 1). When the administration of the alkaloid started at the age of 5 days, decreased levels of CD4⫹, CD8⫹, and Ig⫹ cells were determined. This result is not surprising as the precursor cells might express higher susceptibility to alkaloid’s action than the already differentiated cell clones in mature mice. In Candida infected mice CD4⫹ population was suppressed contrary to the elevated CD8⫹ and Ig⫹ cell levels. Oxoglaucine prevented Candida-induced increase of these populations. Evidently, the alkaloid is able to influence not only the primary immune response but it can affect the late specific response to the yeast. One of the main cytokines needed for the early
N. Ivanovska and M. Hristova / Diagnostic Microbiology and Infectious Disease 38 (2000) 17–20
19
Fig. 1. Effect of oxoglaucine on serum TNF-␣ level of mice with C. albicans infection or adjuvant arthritis. Data are means ⫾SEM from three determinations of individual sera; n ⫽ 5 animals for each point; **p ⬍ 0.01, ***p ⬍ 0.001 by Student’s t-test calculated to the infected group.
protective response against Candida evasion is TNF-␣. The results show that the course of Candida infection is attended with high serum TNF-␣ level with maximum at first day after inoculation. The pretreatment with oxoglaucine in 10 days before the inoculation suppressed to some extent serum TNF-␣ accumulation at first 5 days of infection (Fig. 1). This downregulation of TNF-␣ most probably prevented the pathogen elimination at the initial period of infection. TNF-␣ is also one of the cytokines related to the development of rheumatoid arthritis (Kollias et al. 1999). Different mode of oxoglaucine action was registered in adjuvant arthritis. The alkaloid failed to change the TNF-␣ accumulation until day 14. At the late period it strongly reduced TNF-␣ level, contrary to the second elevation noted in the arthritic group. These different patterns in both processes pointed that various mechanisms might be triggered in each case and it was of interest to determine the consequences of oxoglaucine application in arthritic state in relation to antiinfectious resistance. When arthritic mice were infected at the peak of inflammation (14 day of disease), they were more prone to C. albicans inoculation than the intact mice (Fig. 2). The application of oxoglaucine during 10 days of developing arthritis resulted in protective effect, since the number of survivors was significantly higher as compared to nontreated with oxoglaucine arthritic animals. These results suggest that the ongoing autoimmune process shifts Th1/ Th2 balance to Th2 response. Oxoglaucine possessed suppressive action on host resistance to C. albicans in intact mice but its effect in arthritic animals was beneficial. Thus the alkaloid seems to be perspective for application in au-
toimmune disorders, as its application at the right time might result in less side effects in concern to antiinfectious resistance.
Fig. 2. Effect of oxoglaucine on the outcome of C. albicans infection in mice with adjuvant artritis. Mice (8 w old) were treated i.p. with 10 mg/kg of oxoglaucine from the day of adjuvant injection (day 0) to day 10. Four days later the i.v. inoculation with 1 ⫻ 105 C. albicans cells was done and the infection was monitored daily during 14 days. *p ⬍ 0.001 by 2 test calculated to the group with infection only.
20
N. Ivanovska and M. Hristova / Diagnostic Microbiology and Infectious Disease 38 (2000) 17–20
References Carter, L. L., & Dutton, R. W. (1996). Type 1 and Type 2: A fundamental dichotomy for all T-cell subsets. Curr Opinion Immunol 8, 336 –342. Cenci, E., Mencacci, A., Del Sero, G., Fe` d’Ostiani, C., Mosci, P., Kopf, M., & Romani, L. (1998). IFN- is required for IL-12 responsiveness in mice with Candida albicans infection. J Immunol 161, 3543–3550. Charlton, B., & Lafferty, K. J. (1995). The Th1/Th2 balance in autoimmunity. Curr Opinion Immunol 7, 793–798. Ivanovska, N., Philipov, S., & Georgieva, P. (1997a). Immunopharmacological activity of aporphinoid alkaloid oxoglaucine. Pharmacol Res 35, 267–272. Ivanovska, N., & Philipov, S. (1997b). A low dose immunorestorative effect of aporphinoid alkaloid oxoglaucine on experimentally immunosuppressed and infected mice. Meth Find Exp Clin Pharmacol 19, 579 –583. Ivanovska, N., Hristova, M., & Philipov, S. (2000). Immunosuppression and recovery of drug impaired host resistance against Candida albicans infection by oxoglaucine. Pharmacol Res 41, 99 –105. Kerksiek, K. M., & Pamer, E. G. (1999). T cell responses to bacterial infection. Curr Opinion Immunol 11, 400 – 405.
Kollias, G., Douni, E., Kassiotis, G., & Dimitris, K. (1999). On the role of tumor necrosis factor and receptors in models of multiorgan failure, rheumatoid arthritis, multiple sclerosis and inflammatory bowel disease. Immunol Rev 169, 175–194. Mencacci, A., Cenci, E., Bistoni, F., Bacci, A., Del Sero, G., Montagnoli, C., Fe` d’Ostiani, C., & Romani, L. (1998). Specific and nonspecific immunity to Candida albicans: a lesson from genetically modified animals. Res Immunol 149, 352–361. Romani, L., Mencacci, A., Tonnetti, L., Spaccapelo, R., Cenci, E., Puccetti, P., Wolf, S. F., & Bistoni, F. (1994). Interleukin-12 is both required and prognostic in vivo for T helper type 1 differentiation in murine candidiasis. J Immunol 153, 5157–5175. Romani, L., Mencacci, A., Cenci, E., Spaccapelo, R., Toniatti, C., Puccetti, P., Bistoni, F., & Ploi, V. (1996). Impaired neutrophil response and CD4⫹ T helper cell 1 development in interleukin-6-deficient mice infected with Candida albicans. J Exp Med 183, 1–11. Sakaguchi, S., Toda, M., Asano, M., Itoh, M., Morse, S. S., & Sakaguchi, N. (1996). T cell-mediated maintenance of natural self-tolerance: its breakdown as a possible cause of various autoimmune diseases. J Autoimmun 9, 211–220.