Increased production of chemotactic cytokines and elevated proliferation and expression of intercellular adhesion molecule-1 in rat mesangial cells treated with erythrogenic toxin type B and its precursor isolated from nephritogenic streptococci
Jaimar Rincon1,
Ninoska T. Viera2,
Maritza J. Romero1 and
Jesus A. Mosquera1,
1 Instituto de Investigaciones Clinicas Dr. Americo Negrette, Facultad de Medicina and
2 Instituto de Investigaciones de la Facultad de Odontologia, Facultad de Odontologia, Universidad del Zulia, Maracaibo, Venezuela
 |
Abstract
|
---|
Background. Previous reports have demonstrated the presence of streptococcal erythrogenic toxin type B (ETB) as well as proliferation and expression of adhesion molecules along with leukocyte infiltrations in biopsies from patients with acute post-streptococcal glomerulonephritis (APSGN). The purpose of the present study was to correlate infiltrative and proliferative events with interactions between ETB or its precursor (ETBP) and intrinsic mesangial cells.
Methods. Rat mesangial cells were cultured with ETB or ETBP (50 µg/ml) while measuring production of monocyte chemoattractant protein-1 (MCP-1) and macrophage inflammatory protein-2 (MIP-2) and while examining proliferation and expression of intercellular adhesion molecule-1 (ICAM-1). After 24, 48 and 96 h of incubation, MCP-1 and MIP-2 in culture supernatants were assessed by enzyme-linked immunosorbent assay (ELISA). Cells were assessed for proliferation by incorporation of radioactive thymidine and expression of ICAM-1 was measured by indirect immunofluorescence and by cellular ELISA.
Results. Compared with controls, treatment with either ETBP or ETB significantly increased MCP-1 and MIP-2 levels in mesangial cell cultures. Mesangial cells also showed elevated proliferation at 96 h of culture when treated with streptococcal proteins. Although production of MCP-1 and MIP-2 was not correlated with proliferation, treatment with ETBP resulted in a significant correlation between MCP-1 production and proliferation. Immunofluorescence studies revealed an increased expression of ICAM-1 in ETBP/ETB-treated mesangial cells. In addition, cellular ELISA studies showed increased absorbance in cultures treated with ETBP/ETB. Finally, low serum concentrations in the culture medium potentiated the stimulatory effect of ETB on MCP-1 production.
Conclusions. Our findings, by demonstrating a role for cationic streptococcal ETB or ETBP in the induction of chemotactic molecules as well as the proliferation and expression of adhesion molecules, delineate an additional possible pathway for the pathogenesis of APSGN.
Keywords: erythrogenic toxin; glomerulonephritis; intercellular adhesion molecule-1; macrophage inflammatory protein-2; monocyte chemoattractant protein-1; proliferation
 |
Introduction
|
---|
Acute post-streptococcal glomerulonephritis (APSGN) is a diffuse proliferative endocapillar nephritis that occurs during the convalescent period of group A streptococcal infections [1]. One of the relevant renal histological features of APSGN is an increased glomerular cellularity [24]. Previous studies indicate that the streptococcal erythrogenic toxin type B (ETB) and its precursor (ETBP) could be involved in the pathogenesis of APSGN. In this regard, ETB has been found in renal biopsies from patients with APSGN and APSGN sera react preferentially with these streptococcal proteins [58]. The presence of streptococcal proteins in the renal microenvironment creates possible interactions between these proteins and intrinsic glomerular cells and this interaction may lead to increased glomerular cellularity. In support of this we previously reported increased glomerular and interstitial leukocyte infiltration after in vivo renal perfusion of ETB or ETBP, an effect that may have been mediated by chemotactic and macrophage migration inhibition factor (MIF) activities [9]. We additionally reported that ETB and ETBP have capacities to induce proliferation in human mononuclear leukocytes [10]. On the basis of these observations, combined with findings that infiltration, proliferation and expression of adhesion molecules are involved in the hypercellularity observed during the course of glomerulonephritis [14,11], we studied the effects of ETB and ETBP on the proliferation and production of monocyte chemoattractant protein-1 (MCP-1), macrophage inflammatory protein-2 (MIP-2) and expression of intercellular adhesion molecule-1 (ICAM-1) in cultured mesangial cells. Our findings demonstrated that ETB and ETBP were capable of inducing increased production of MCP-1 and MIP-2 and of upregulating proliferation and increased expression of ICAM-1 by mesangial cells.
 |
Subjects and methods
|
---|
Mesangial cell cultures
Mesangial cells were isolated from the glomeruli of SpragueDawley rats using a differential sieving technique. Briefly, cortices were removed and minced before passage through a series of steel sieves with decreasing pore sizes (200, 150 and 75 µm). Isolated glomeruli were then digested with collagenase. Digested glomeruli were cultured in RPMI 1640 combined with antibiotics (100 U/ml penicillin and 10 µg/ml streptomycin) and 20% fetal bovine serum (FBS; Sigma Chemical Co., St Louis, MO) until mesangial cells reached confluence. The cells were passaged using trypsin/EDTA and cells between passages 3 and 6 were used for experiments. Cells were characterized and the homogeneity of mesangial cells was assessed by identifying typical morphology and patterns of staining with antibodies against smooth muscle cell-specific actin, Thy 1.1, factor VIII antigen, leukocyte common antigen and by the presence of hillocks [12]. Mesangial cells were cultured until subconfluence (70%) and treated with 50 µg/ml cationic ETB or ETBP (final concentration) for 24, 48 and 96 h at 37°C with 5% CO2 in the supplemented media as previously described. Blocking experiments were performed by incubation of ETB-treated mesangial cell cultures with a rabbit anti-ETB polyclonal antibody (100 µg/ml). To determine the effect of low level FBS on the production of chemotactic cytokines, mesangial cell cultures were incubated for 96 h with or without ETB in RPMI, penicillin/streptomycin and 0.5% FBS. Control experiments used cellular cultures without streptococcal proteins. The cationic streptococcal protein concentration was chosen from previous investigations [12,14] that reported doses ranging from 20 to 500 µg of cationic antigens to induce in situ immune complex glomerulonephritis in rats and from our demonstration of an optimal proliferative-inducer effect of ETB and ETBP at a dose of 50 µg/ml in human mononuclear leukocytes [10]. After incubation, cells or supernatants were tested for production of MCP-1 and MIP-2 and proliferation and expression of ICAM-1.
Isolation of ETBP and ETB
The streptococcal ETB and ETBP were kindly donated by Drs Arnold Vogt and Stephen Batsford. To isolate these, we used two ß-haemolytic group A streptococcal strains isolated from patients with APSGN. The bacteria were kept on blood agar at 4°C and pre-cultured in chemically defined medium (CDM), pH 6.9 at 37°C for 6 h. Following this, 20 ml of bacterial culture were added to 180 ml of fresh CDM and the mixture was cultured until it was cloudy (
6 h). Two-hundred ml of the pre-culture were added to the main culture medium (800 ml CDM and 1000 ml Todd-Hewitt, 1% glucose, pH 6.9). Bacteria were cultured for several hours and pH was maintained by adding of 0.5 N NaOH with 50% glucose. Then, pH was reduced to 5.9 and was maintained at this level for 12 h with a 0.5 N NaOH solution containing 50% glucose. The culture was centrifuged (10 800 gx15 min) and the supernatant was sterile filtered, concentrated to a factor of 1000 in an Amicon system using an YM-5 membrane and finally run on a Mono S cation exchange column (Pharmacia HR 5/5) in a fast-performance liquid chromatography system. The system conditions included: pH 6.0, flow rate 1 ml/min and a buffer gradient of 3.5250 mM MES. The ETBP eluted before ETB and the fractions were tested in SDSPAGE under non-reducing conditions. Then, streptococcal antigens were run in isoelectric focusing to determine pI. Under these conditions, ETBP was 44 kDa, pI 8.2 and ETB was 30 kDa, pI 9.0. ETBP was exposed to iodoacetamide to avoid its autocatalytic transformation into ETB. A Limulus amebocyte lysate assay revealed that streptococcal proteins solutions were free of endotoxin.
Detection of MCP-1 and MIP-2 in culture supernatants
Supernatants from mesangial cell cultures in the absence or presence of streptococcal proteins (ETB/ETBP) were harvested at 24, 48 and 96 h and were centrifuged and stored at 80°C until use. MCP-1 and MIP-2 secreted into the supernatant were quantified using a commercially available enzyme-linked immunosorbent assay (ELISA) kit (Biosource International Inc., CA, USA) according to the manufacturer's protocol. Amounts of MCP-1 and MIP-2 were expressed as pg per 10 µg cellular culture protein. For the protein assay, mesangial cell cultures were lysed for 10 min at 4°C in a lysis buffer (20 mM Tris, pH 8.0, 137 mM NaCl and 1% Triton X-100). The lysates were collected and protein content was determined by the Lowry protein assay.
Determination of ICAM-1 protein in mesangial cells
To examine the expression of ICAM-1 in mesangial cell cultures in the presence or absence of streptococcal proteins, mesangial cells were planted on 8-well plastic chamber slides (Nunc, Roskilde, Denmark) and incubated until subconfluence. Treated and untreated cultures were washed with phosphate-buffered saline (PBS) and fixed with 4% paraformaldehyde for 10 min. After washing, cells were reacted with monoclonal antibody against rat ICAM-1 (Seikagaku Co., Tokyo, Japan) for 30 min and thereafter, a fluorescein isothiocyanate-conjugated goat anti-mouse IgG was used (Accurate Chemical Co., NY, USA) to localize the first antibody. As a negative control mesangial cells were incubated, as previously described, with an irrelevant monoclonal antibody. ICAM-1 expression was also measured by ELISA. In brief, mesangial cells were cultured in flat bottom 96-well plates. Cells were incubated in the absence or presence of ETB or ETBP and incubated for 24, 48 and 96 h. After incubation, the cells were washed with PBS and fixed with 4% paraformaldehyde for 15 min. Cells were then washed with PBS and blocked with 3% ovalbumin in PBS. The cells were then incubated with an anti-rat ICAM-1 monoclonal antibody (1:100). After incubation for 30 min at room temperature, cells were washed with 3% ovalbumin and incubated with a goat anti-mouse IgGhorseradish peroxidase conjugated antibody (1:200; Pierce, IL, USA) for an additional 30 min. After washing, the ELISA was developed using 3,3',5'5-tetramethylbenzidine (Sigma Chemical Co., St Louis, MO, USA) in citrate buffer containing 0.006% hydrogen peroxide. Absorbance was measured at 450 nm using a Benhmark model microplate reader (Biorad, CA, USA).
Proliferation assay
Subconfluent mesangial cell monolayers in 96-well flat-bottom tissue culture plates with RPMI 1640 supplemented with 20% FBS were incubated with 50 µg/ml (final concentration) ETB or ETBP for 24, 48 and 96 h at 37°C in 5% CO2 atmosphere. Control cultures received supplemented medium without streptococcal proteins. Eighteen hours before cell harvest, cells were pulsed with 0.5 µCi/well of 3H-thymidine (NEN Research Products, DuPont, MA, USA). Adherent cells were trypsinized and harvested on individual filter discs, dried and placed in 3 ml scintillation fluid. Radioactivity was counted in a beta scintillation counter. Control and experimental cultures were performed in triplicate and data were expressed as counts per minute (CPM).
Statistical analysis
Results in the groups are shown as means±SEM and represent data from three to four individual experiments. Comparisons between groups were performed by ANOVA and associations between variables were analysed by linear correlation (Pearson). Two-tailed P-values of <0.05 were considered statistically significant.
 |
Results
|
---|
Induction of chemokines and proliferation of cultured mesangial cells caused by ETB and ETBP
The incubation of mesangial cells with ETB or ETBP led to increased amounts of MCP-1 and MIP-2 in the supernatant cultures. The production of both chemokines was already demonstrable at 24 h and increased over time (Figures 1
and 2
). Mesangial cells incubated in the absence of streptococcal proteins had basal levels of MCP-1 and MIP-2 at 24 h; there was no significant spontaneous chemokine production at 48 and 96 h of culture (Figures 1
and 2
). Treatment of ETB mesangial cell cultures with an anti-ETB polyclonal antibody abolished the stimulatory effect of ETB on MCP-1 production (Figure 3
). ETB-treated mesangial cell cultures under low serum conditions (0.5% FBS) showed increased concentrations of MCP-1 in the supernatants compared with 20% FBS cultures (Figure 4
).

View larger version (28K):
[in this window]
[in a new window]
|
Fig. 1. Effect of ETBP and ETB on the production of MCP-1 by cultured mesangial cells. Mesangial cells were cultured with 50 µg/ml ETBP or ETB for 24, 48 and 96 h. Control cells were cultured in absence of streptococcal proteins. MCP-1 from culture supernatants was determined by ELISA. Values are expressed as means±SE from four independent experiments, each conduced in triplicate. *P<0.05.
|
|

View larger version (41K):
[in this window]
[in a new window]
|
Fig. 2. Effect of ETBP and ETB on the production of MIP-2 in cultured mesangial cells. Mesangial cells were incubated in the absence or presence of 50 µg/ml ETBP or ETB for 24, 48 and 96 h. Concentrations of MIP-2 in the culture supernatants were obtained by ELISA. Values are expressed as means±SE from four independent experiments, each conduced in triplicate. *P<0.05.
|
|

View larger version (32K):
[in this window]
[in a new window]
|
Fig. 3. Loss of stimulatory effect of ETB on MCP-1 after incubation of ETB mesangial cell cultures with an anti-ETB antibody. Mesangial cells were incubated with ETB (50 µg/ml) and anti-ETB antibody (100 µg/ml) for 96 h and MCP-1 in the supernatant was determined by ELISA.
|
|

View larger version (30K):
[in this window]
[in a new window]
|
Fig. 4. MCP-1 production in ETB-treated mesangial cell cultures with different serum concentrations. Mesangial cells were cultured with ETB (50 µg/ml) in 20% or 0.5% FBS for 96 h. MCP-1 levels were significantly increased in 0.5% FBS cultures compared with 20% cultures. *P<0.01.
|
|
Proliferation of mesangial cells was upregulated by ETB and ETBP. In the presence of streptococcal proteins, mesangial cells reached a significant uptake of 3H-thymidine at 96 h of culture (Figure 5
). Figure 6
shows the kinetic curves of proliferation and production of MCP-1 and MIP-2, in which MCP-1 production was associated with proliferation in mesangial cell cultures treated with ETBP. There was no correlation between proliferation and chemokine production by ETB and by ETBP, except in ETBP-treated mesangial cell cultures, in which MCP-1 production was significantly correlated with proliferation (r=0.803, P=0.0052).

View larger version (14K):
[in this window]
[in a new window]
|
Fig. 5. Proliferative effect of ETBP and ETB on cultured mesangial cells. Mesangial cell proliferation was upregulated by ETB and ETBP. The presence of streptococcal proteins caused increased uptake of 3H-thymidine by cellular cultures at 96 h of culture. Control experiments used unstimulated cultures. Values are expressed as means±SE from four independent experiments, each conduced in triplicate. *P<0.05.
|
|

View larger version (20K):
[in this window]
[in a new window]
|
Fig. 6. Kinetics of MCP-1 and MIP-2 as well as mesangial cell culture proliferation in the presence or absence of streptococcal proteins. Although MCP-1 production was associated with proliferation in mesangial cell cultures treated with ETBP (A), there was no association with MIP-2 production or when ETB was used to stimulate the cultures (A and B). Unstimulated cultures are shown in (C).
|
|
Effect of ETBP and ETB on ICAM-1 expression in cultured mesangial cell
To investigate whether streptococcal proteins could also stimulate ICAM-1 expression in mesangial cells, cellular cultures were incubated with 50 µg/ml of ETBP or ETB for 24, 48 and 96 h. Although basal expression of ICAM-1 was found in untreated cultures, stimulation with ETB or ETBP increased expression of ICAM-1 (Figure 7
). These observations were confirmed by cellular ELISA (Figure 8
).

View larger version (143K):
[in this window]
[in a new window]
|
Fig. 7. Indirect immunofluorescence staining for ICAM-1 in ETB-treated mesangial cells at 96 h of culture. Magnification: x400.
|
|

View larger version (38K):
[in this window]
[in a new window]
|
Fig. 8. Time course of ICAM-1 expression in cultured mesangial cells. As described in the Subjects and Methods, untreated and ETBP- or ETB-treated mesangial cell cultures were fixed with paraformaldehyde and reacted with a monoclonal antibody against rat ICAM-1 in a cellular ELISA. Values are expressed as means±SE from three independent experiments, each conduced in triplicate. *P<0.01.
|
|
 |
Discussion
|
---|
Overall renal histologic patterns in APSGN consistently show moderate to marked hypercellularity that is primarily due to proliferation of mesangial and endothelial cells. In the very early stage there is increased non-glomerular cell infiltration, consisting of polymorphonuclear leukocytes and monocyte/macrophage cells [14]. The renal tissue inflammation observed in APSGN is generally believed to result from actions of anaphylatoxins generated by the classic complement pathway [11,15,16]. However, Streptococcus pyogenes produces several extracellular proteins, including ETB, that may be involved in APSGN-stimulated renal pathology. The role of streptococcal ETB and ETBP in APSGN has been documented by the increased reactivity to ETB and ETBP in APSGN sera and by the presence of ETB in kidney biopsies from APSGN patients [58]. In this regard, our data suggesting that ETB/ETBP may be involved in the renal hypercellularity after in vivo renal perfusion indicate that ETB and ETBP are potentially nephritogenic by themselves. This effect could be mediated by chemotaxis mechanisms and by MIF activity [9]. ETB/ETBP may also induce proliferation of human mononuclear leukocytes [10], an important cellular component during APSGN [14].
Our experiments were designed to elucidate whether the interaction between ETB/ETBP and mesangial cells is related to the renal hypercellularity observed in APSGN. This study provided evidence that ETB/ETBP from nephritogenic streptococci can induce mesangial cells to increase the production of MIP-2 and MCP-1. MIP-2 is an alpha chemokine and a major neutrophil chemoattractant contributing to influx of neutrophils in several forms of glomerulonephritis [1720]. MCP-1 is a beta chemokine that is very important in recruiting and activating monocyte/macrophages during the course of certain types of glomerulonephritis [19,2124]. In addition to the chemokine-inducer effects of ETB/ETBP, these streptococcal products were capable of inducing proliferation of cultured mesangial cells. Both proliferative and chemotactic mechanisms are involved in the hypercellularity observed during the course of several experimental and human forms of nephritis [1,3,11,25]. The presence of streptococcal proteins in the mesangial microenvironment [57,9] indicates a possible interaction between ETB/ETBP and intrinsic mesangial cells during APSGN, leading to increased expression of MCP-1 and MIP-2 and further influx of neutrophils and monocytes into renal tissues. These events along with the increased proliferation of mesangial cells may play a role in the pathogenesis of APSGN. The increased influx of monocytes and neutrophils induced by MCP-1 and MIP-2 could promote increased production of macrophage and neutrophil inflammatory cytokines and oxygen reactive species to cause further tissue damage [26,1720]. In turn, certain macrophage products may induce increased expression of MCP-1 by mesangial cells in a paracrine manner [2729]. In general, the chemokine inducer and the proliferative effects of ETB/ETBP were not correlated, suggesting that the increased production of chemokines was not only due to the proliferation of mesangial cells. In addition, the expression of chemokine concentration per cellular protein content showed increased production of chemokines. However, MCP-1 production and proliferation were correlated when mesangial cells were treated with ETBP. We have no clear explanation for this effect, but activation of a common transcription factor for both processes by streptococcal products could be involved [3033]. Unexpectedly, stimulation of mesangial cells with ETB during low serum concentrations (0.5% FBS) caused a greater production of MCP-1 than during 20% FBS, suggesting the presence of inhibitory factors in FBS. However, histological examination revealed that low serum cultures had morphological cellular alterations and cellular detachment. In addition, low serum cultures had decreased protein content in the cellular lysate (20% serum: 114±27.05 µg/ml; 0.5% serum: 52.21±21.88 µg/ml). We therefore decided to perform the remainder of the experiments under 20% serum conditions.
In the present study, we also documented increased expression in ICAM-1 on ETB/ETBP-treated cultured mesangial cells. This finding suggests that resident mesangial cells may increase ICAM-1 expression in APSGN following interaction with streptococcal proteins and may partly explain the increased expression of glomerular ICAM-1 found in early biopsies from patients with APSGN [34]. Since macrophages accumulate in the mesangium during APSGN, the increased expression of mesangial ICAM-1 may potentiate the proliferative effect of direct macrophagemesangial cell contact on mesangial cells [35]. Increased ICAM-1 expression on the mesangial cell surface may also promote the recruitment of leukocytes in the mesangium by interaction with its LFA-1 ligand that may be relevant in mesangial hypercellularity. In support of this, increased expression of glomerular LFA-1 positive cells and increased expression of glomerular ICAM-1 have been reported in APSGN [34]. Previous work has shown the importance of ICAM-1 in the antigen presentation function in mesangial cells [36]. Presentation of specific antigens by mesangial cells to T cells that are initially engaged by ICAM-1 may lead to a rapid T-cell activation and production of cytokines that could increase major histocompatibility complex class II antigens on mesangial cells to regulate the specificity of the immune response and amplify its intensity [3740]. Since both the presence of streptococcal antigens and increased expression of ICAM-1 and T lymphocytes have been demonstrated in the renal mesangium of patients with APSGN [2,3,57,9,34], it is possible that ETB/ETBP may induce a local immune response that contributes to the mesangial expansion observed in APSGN.
In conclusion, we found that ETB/ETBP from nephritogenic streptococci may induce increased production of chemokines and cause increased proliferation and expression of ICAM-1 in mesangial cells, which provide mechanisms that may partly explain the inflammatory process observed during the nephritis. These effects of streptococcal proteins on mesangial cells, along with other inflammatory mechanisms, underline their potential pathogenic role in APSGN.
 |
Acknowledgments
|
---|
The authors thank Drs Arnold Vogt and Stephen Batsford for their generous gift of ETBP, ETB and antibodies against streptococcal proteins. This work was supported by the Instituto de Investigaciones Clinicas Dr Americo Negrette. These data were presented at the XXXVI Congress of the European Dialysis and Transplant Association, Vienna, Austria, June 2001 and were published in abstract form (ERAEDTA Abstracts, pp. 4546, 2001).
Conflict of interest statement. None declared.
 |
Notes
|
---|
Correspondence and offprint requests to: Jesus A. Mosquera, MD, Apartado Postal 1151, Maracaibo 4001-A, Zulia, Venezuela. Email: mosquera99{at}hotmail.com 
 |
References
|
---|
- Rammelkamp CH. Acute poststreptococcal glomerulonephritis. In: Read SE, Zabriskie JB, eds. Streptococcal Diseases and the Immune Response. Academic Press, New York, NY:1980; 4357
- Parra G, Plat JL, Falk RJ, Rodriguez-Iturbe B, Michael AF. Cell populations and membrane attack complex in glomeruli of patients with post-streptococcal glomerulonephritis: identification using monoclonal antibodies by indirect immunofluorescence. Clin Immunol Immunopathol 1984; 33:324332[CrossRef][ISI][Medline]
- Hooke DH, Gee DC, Atkins RC. Leukocyte analysis using monoclonal antibodies in human glomerulonephritis. Kidney Int 1987; 31:964972[ISI][Medline]
- Soto H, Mosquera J, Rodríguez-Iturbe B, Henriquez-La Roche C, Pinto A. Apoptosis in proliferative glomerulonephritis: decreased apoptosis expression in lupus nephritis. Nephrol Dial Transplant 1997; 12:273280[Abstract]
- Vogt A, Batsford S, Rodriguez-Iturbe B, Garcia R. Cationic antigens in poststreptococcal glomerulonephritis. Clin Nephrol 1983; 20:271279[ISI][Medline]
- Vogt A, Schmiedeke T, Stockl F, Sugisaki Y, Mertz A, Batsford S. The role of cationic proteins in the pathogenesis of immune complex glomerulonephritis. Nephrol Dial Transplant 1990; 5 [Suppl 1]:69[ISI][Medline]
- Cu GA, Mezzano S, Bannan JD, Zabriskie JB. Immunohistochemical and serological evidence for the role of streptococcal proteinase in acute post-streptococcal glomerulonephritis. Kidney Int 1998; 54:819826[CrossRef][ISI][Medline]
- Parra G, Rodriguez-Iturbe B, Batsford S et al. Antibody to streptococcal zymogen in the serum of patients with acute glomerulonephritis: a multicentric study. Kidney Int 1998; 54:509517[CrossRef][ISI][Medline]
- Romero M, Mosquera J, Novo E, Fernandez L, Parra G. Erythrogenic toxin type B and its precursor isolated from nephritogenic streptococci induce leukocyte infiltration in normal rat kidneys. Nephrol Dial Transplant 1999; 14:18671874[Abstract/Free Full Text]
- Viera NT, Romero MJ, Montero MK, Rincon J, Mosquera JA. Streptococcal erythrogenic toxin B induces apoptosis and proliferation in human leukocytes. Kidney Int 2001; 59:950958[CrossRef][ISI][Medline]
- Courser WG. Mechanisms of glomerular injury in immune complex disease. Kidney Int 1985; 28:569583[ISI][Medline]
- Sterzel RB, Lovett DH, Foellner HG, Perfetto M, Biemesderfer D, Kashgarian M. Mesangial cell hillocks. Nodular foci of exaggerated growth of cells and matrix in prolonged culture. Am J Pathol 1986; 125:130140[Abstract]
- Oite T, Batsford SR, Mihatsch MJ, Takamiya H, Vogt A. Quantitative studies of in situ immune complex glomerulonephritis in the rat induced by planted cationized antigen. J Exp Med 1982; 155:460474[Abstract/Free Full Text]
- Yousif Y, Okada K, Batsford S, Vogt A. Induction of glomerulonephritis in rat with staphylococcal phosphatase: new aspects in post-infectious ICGN. Kidney Int 1996; 50:290297[ISI][Medline]
- Liszewski MR, Farries TC, Lublin DM, Rooney IA, Atkinson JP. Control of complement system. Adv Immunol 1996; 61:202283
- Michael AF, Kim Y. Pathogenesis of acute post-streptococcal glomerulonephritis. In: Reed SE, Zabriskie JB, eds. Streptococcal Disease and the Immune Response. Academic Press, New York, NY:1980;7992
- Walpen S, Beck KF, Schaefer L et al. Nitric oxide induces MIP-2 transcription in rat renal mesangial cells and in a rat model of glomerulonephritis. FASEB J 2001; 15:571573[Abstract/Free Full Text]
- Feng L, Xia Y, Yoshimura T, Wilson CB. Modulation of neutrophil influx in glomerulonephritis in the rat with anti-macrophage inflammatory protein-2 (MIP-2) antibody. J Clin Invest 1995; 95:10091017[ISI][Medline]
- Tam FW, Karkar AM, Smith J et al. Differential expression of macrophage inflammatory protein-2 and monocyte chemoattractant protein-1 in experimental glomerulonephritis. Kidney Int 1996; 49:715721[ISI][Medline]
- Tang WW, Yin S, Wittwer AJ, Qi M. Chemokine gene expression in anti-glomerular basement membrane antibody glomerulonephritis. Am J Physiol 1995; 269:F323F330[ISI][Medline]
- Prodjosudjadi W, Gerritsma JSJ, van Es LA, Daha MR, Bruijn JA. Monocyte chemoattractant protein-1 in normal and diseased human kidneys: an immunohistochemical analysis. Clin Nephrol 1995; 44:148155[ISI][Medline]
- Sekiguchi M, Natori Y, Iyonaga K, Takeya M, Natory Y. Expression of monocyte chemoattractant protein-1 in experimental crescentic glomerulonephritis in rats. J Lab Clin Med 1997; 129:239244[CrossRef][ISI][Medline]
- Grandaliano G, Gesualdo L, Ranieri E et al. Monocyte chemotactic peptide-1 expression in acute and chronic human nephritides: a pathogenetic role in interstitial monocytes recruitment. J Am Soc Nephrol 1996; 7:906913[Abstract]
- Stahl RAK, Thaiss F, Disser M, Helmchen U, Hora K, Schlöndorff D. Increased expression of monocyte chemoattractant protein-1 in anti-thymocyte antibody-induced glomerulonephritis. Kidney Int 1993; 44:10361047[ISI][Medline]
- Monga G, Mazzuco G, Belgiojoso GB, Busnach G. Monocyte infiltration and glomerular hypercellularity in human acute and persistent glomerulonephritis; light and electron microscopic, immunofluorescence, and histochemical investigation on twenty eight cases. Lab Invest 1981; 44:381387[ISI][Medline]
- Cattell V. Macrophages in acute glomerular inflammation. Kidney Int 1994; 45:945952[ISI][Medline]
- Ishikawa Y, Sugiyama H, Stylianou E, Kitamura M. Bioflavonoid quercetin inhibits interleukin-1 induced transcriptional expression of monocyte chemoattractant protein-1 in glomerular cells via suppression of nuclear factor-kappa B. J Am Soc Nephrol 1999; 10:22902296[Abstract/Free Full Text]
- Rovin BH, Tan LC. Role of protein kinase pathways in IL-1 induced chemoattractant expression by human mesangial cells. Kidney Int 1994; 46:10591068[ISI][Medline]
- Prodjosudjadi W, Gerritsma JSJ, Klar-Mohamad N et al. Production and cytokine-mediated regulation of monocyte chemoattractant protein-1 by human proximal tubular epithelial cells. Kidney Int 1995; 48:14771486[ISI][Medline]
- Landry DB, Couper LL, Bryant SR, Lindner V. Activation of the NF-kappa B and I kappa B system in smooth muscle cells after rat arterial injury. Induction of vascular cell adhesion molecule-1 and monocyte chemoattractant protein-1. Am J Pathol 1997; 151:10851095[Abstract]
- Goebeler M, Gillitzer R, Kilian K et al. Multiple signaling pathways regulate NF-kappa B-dependent transcription of the monocyte chemoattractant protein-1 gene in primary endothelial cells. Blood 2001; 97:4655[Abstract/Free Full Text]
- Joyce D, Albanese C, Steer J, Fu M, Bouzahzah B, Pestell RG. NF-kappa B and cell-cycle regulation: cyclin connection. Cytokine Growth Factor Rev 2001; 12:7390[CrossRef][ISI][Medline]
- Miettinen M, Lehtonen A, Julkunen I, Matikainen S. Lactobacilli and streptococci activate NF-kappa B and STAT signaling pathways in human macrophages. J Immunol 2000; 164:37333740[Abstract/Free Full Text]
- Parra G, Romero M, Henriquez-La Roche C, Pineda R, Rodriguez-Iturbe B. Expression of adhesion molecules in post-streptococcal glomerulonephritis. Nephrol Dial Transplant 1994; 9:14121417[Abstract]
- Kasai S, Mori T, Komiyama A, Ito N, Shigematsu H. The importance of cellular adhesion for mesangial cell proliferation in serum sickness nephritis of the rat: a co-culture study of glomerular macrophages and mesangial cells. Pathol Int 1994; 44:413419[ISI][Medline]
- Brennan DC, Jevnikar AM, Takei F, Reubin-Kelley VE. Mesangial cell accessory functions: mediation by intercellular adhesion molecule-1. Kidney Int 1990; 38:10391046[ISI][Medline]
- Radeke HH, Resch K. The inflammatory function of renal glomerular mesangial cells and their interaction with the cellular immune system. Clin Invest 1992; 70:825842[ISI][Medline]
- Radeke HH, Emmendorffer A, Uciechowski P, von der Ohe J, Schwinzer B, Resch K. Activation of autoreactive T-lymphocytes by cultured syngenic glomerular mesangial cells. Kidney Int 1994; 45:763774[ISI][Medline]
- Lub M, van Kooyk Y, Figdor CG. Ins and outs of LFA-1. Immunol Today 1995; 16:479483[CrossRef][ISI][Medline]
- Mosquera J. Interferon gamma induces class II antigen expression on cultured rat mesangial smooth muscle cells. Clin Immunol Immunopathol 1989; 53:341345[CrossRef][ISI][Medline]
Received for publication: 29. 1.02
Accepted in revised form: 2.12.02