Streptococcal erythrogenic toxin B induces apoptosis and proliferation in human leukocytes

Ninoska T. Viera, Maritza Josefina Romero Lucas, Mary K. Montero, Jaimar Rincon, Jesus A. Mosquera

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Background. Previous reports have shown the presence of erythrogenic toxin type B (ETB), apoptosis, proliferation, and leukocyte infiltration in biopsies from patients with acute poststreptococcal glomerulonephritis (APSGN). Methods. Attempting to correlate the apoptotic and proliferative events with the interaction of ETB or its precursor (ETBP) with leukocytes, mononuclear leukocytes from 12 healthy subjects were cultured with ETB or ETBP to analyze the levels of apoptosis, proliferation, expression of modulatory apoptosis gene products, and oxidative metabolism. After four days of incubation, cells were assessed for apoptosis by morphological criteria, annexin V assay, and terminal deoxy transferase uridine triphosphate nick end-labeling (TUNEL) assay. The expression of regulatory apoptosis genes was assessed by relevant monoclonal antibodies; proliferation was by incorporation of radioactive thymidine; and oxidative metabolism was by oxidation of 2′,7′-dichlorofuorescein diacetate to 2′,7′-dichlorofuorescein. Neutralization of Fas-L and cysteine protease activity of ETB were performed by incubation of ETB-treated leukocyte cultures with anti-human Fas-L mAb or with E64, respectively. Results. Elevated levels of apoptosis in ETBP/ETB-treated leukocytes were found when compared with controls: morphological criteria (P < 0.01), Annexin V (control, 5.01 ± 0.61; ETBP, 10.60 ± 1.98%, P = 0.0005), and TUNEL (control, 12.5 ± 2.6; ETBP, 20.56 ± 3.06%, P = 0.001; ETB, 30.69 ± 5.05%, P = 0.001). Increased expression of apoptosis was accompanied by increased expression of Fas (control, 20.15 ± 5.28; ETBP, 43.51 ± 5.6%, P = 0.03; ETB, 47.16 ± 5.54%, P = 0.01), Fas ligand (control, 5.64 ± 2.38; ETBP, 11.66 ± 3.65%, P = 0.04; ETB, 16.39 ± 5.05%, P = 0.02) and p53 products (control, 9.22 ± 3.44; ETBP, 22.82 ± 5.72%, P = 0.01; ETB, 24.60 ± 5.20%, P = 0.01). Treatment of ETB-leukocyte cultures with anti-human Fas-L exhibited 2.2-fold lower apoptosis expression. Treatment with E64 significantly abrogated the apoptotic effect of ETB. There was no increment on leukocyte oxidative metabolism. Mononuclear leukocytes also showed elevated levels of proliferation when treated with different concentrations (from 50 to 6.2 μg/mL) of streptococcal proteins (Stimulation index ranging: ETBP, 5.6 ± 1.9 to 6.4 ± 1.9; ETB, 9.9 ± 2.8 to 13.9 ± 3.8). Conclusions. These results delineate an additional pathway for the pathogenesis of APSGN related to the role of cationic streptococcal ETB or ETBP on the induction of apoptosis and proliferation during the course of the disease.

Original languageEnglish (US)
Pages (from-to)950-958
Number of pages9
JournalKidney International
Volume59
Issue number3
DOIs
StatePublished - Jan 1 2001
Externally publishedYes

Fingerprint

Leukocytes
Apoptosis
Mononuclear Leukocytes
Uridine Triphosphate
Annexin A5
Transferases
Glomerulonephritis
erythrogenic toxin
Fas Ligand Protein
Cysteine Proteases
Regulator Genes
Thymidine
Cysteine
Healthy Volunteers
Monoclonal Antibodies
Biopsy

Keywords

  • Cell death
  • DNA fragmentation
  • Glomerulonephritis
  • Phosphatidylserine
  • Streptococcal proteinase

ASJC Scopus subject areas

  • Nephrology

Cite this

Streptococcal erythrogenic toxin B induces apoptosis and proliferation in human leukocytes. / Viera, Ninoska T.; Romero Lucas, Maritza Josefina; Montero, Mary K.; Rincon, Jaimar; Mosquera, Jesus A.

In: Kidney International, Vol. 59, No. 3, 01.01.2001, p. 950-958.

Research output: Contribution to journalArticle

Viera, Ninoska T. ; Romero Lucas, Maritza Josefina ; Montero, Mary K. ; Rincon, Jaimar ; Mosquera, Jesus A. / Streptococcal erythrogenic toxin B induces apoptosis and proliferation in human leukocytes. In: Kidney International. 2001 ; Vol. 59, No. 3. pp. 950-958.
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title = "Streptococcal erythrogenic toxin B induces apoptosis and proliferation in human leukocytes",
abstract = "Background. Previous reports have shown the presence of erythrogenic toxin type B (ETB), apoptosis, proliferation, and leukocyte infiltration in biopsies from patients with acute poststreptococcal glomerulonephritis (APSGN). Methods. Attempting to correlate the apoptotic and proliferative events with the interaction of ETB or its precursor (ETBP) with leukocytes, mononuclear leukocytes from 12 healthy subjects were cultured with ETB or ETBP to analyze the levels of apoptosis, proliferation, expression of modulatory apoptosis gene products, and oxidative metabolism. After four days of incubation, cells were assessed for apoptosis by morphological criteria, annexin V assay, and terminal deoxy transferase uridine triphosphate nick end-labeling (TUNEL) assay. The expression of regulatory apoptosis genes was assessed by relevant monoclonal antibodies; proliferation was by incorporation of radioactive thymidine; and oxidative metabolism was by oxidation of 2′,7′-dichlorofuorescein diacetate to 2′,7′-dichlorofuorescein. Neutralization of Fas-L and cysteine protease activity of ETB were performed by incubation of ETB-treated leukocyte cultures with anti-human Fas-L mAb or with E64, respectively. Results. Elevated levels of apoptosis in ETBP/ETB-treated leukocytes were found when compared with controls: morphological criteria (P < 0.01), Annexin V (control, 5.01 ± 0.61; ETBP, 10.60 ± 1.98{\%}, P = 0.0005), and TUNEL (control, 12.5 ± 2.6; ETBP, 20.56 ± 3.06{\%}, P = 0.001; ETB, 30.69 ± 5.05{\%}, P = 0.001). Increased expression of apoptosis was accompanied by increased expression of Fas (control, 20.15 ± 5.28; ETBP, 43.51 ± 5.6{\%}, P = 0.03; ETB, 47.16 ± 5.54{\%}, P = 0.01), Fas ligand (control, 5.64 ± 2.38; ETBP, 11.66 ± 3.65{\%}, P = 0.04; ETB, 16.39 ± 5.05{\%}, P = 0.02) and p53 products (control, 9.22 ± 3.44; ETBP, 22.82 ± 5.72{\%}, P = 0.01; ETB, 24.60 ± 5.20{\%}, P = 0.01). Treatment of ETB-leukocyte cultures with anti-human Fas-L exhibited 2.2-fold lower apoptosis expression. Treatment with E64 significantly abrogated the apoptotic effect of ETB. There was no increment on leukocyte oxidative metabolism. Mononuclear leukocytes also showed elevated levels of proliferation when treated with different concentrations (from 50 to 6.2 μg/mL) of streptococcal proteins (Stimulation index ranging: ETBP, 5.6 ± 1.9 to 6.4 ± 1.9; ETB, 9.9 ± 2.8 to 13.9 ± 3.8). Conclusions. These results delineate an additional pathway for the pathogenesis of APSGN related to the role of cationic streptococcal ETB or ETBP on the induction of apoptosis and proliferation during the course of the disease.",
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TY - JOUR

T1 - Streptococcal erythrogenic toxin B induces apoptosis and proliferation in human leukocytes

AU - Viera, Ninoska T.

AU - Romero Lucas, Maritza Josefina

AU - Montero, Mary K.

AU - Rincon, Jaimar

AU - Mosquera, Jesus A.

PY - 2001/1/1

Y1 - 2001/1/1

N2 - Background. Previous reports have shown the presence of erythrogenic toxin type B (ETB), apoptosis, proliferation, and leukocyte infiltration in biopsies from patients with acute poststreptococcal glomerulonephritis (APSGN). Methods. Attempting to correlate the apoptotic and proliferative events with the interaction of ETB or its precursor (ETBP) with leukocytes, mononuclear leukocytes from 12 healthy subjects were cultured with ETB or ETBP to analyze the levels of apoptosis, proliferation, expression of modulatory apoptosis gene products, and oxidative metabolism. After four days of incubation, cells were assessed for apoptosis by morphological criteria, annexin V assay, and terminal deoxy transferase uridine triphosphate nick end-labeling (TUNEL) assay. The expression of regulatory apoptosis genes was assessed by relevant monoclonal antibodies; proliferation was by incorporation of radioactive thymidine; and oxidative metabolism was by oxidation of 2′,7′-dichlorofuorescein diacetate to 2′,7′-dichlorofuorescein. Neutralization of Fas-L and cysteine protease activity of ETB were performed by incubation of ETB-treated leukocyte cultures with anti-human Fas-L mAb or with E64, respectively. Results. Elevated levels of apoptosis in ETBP/ETB-treated leukocytes were found when compared with controls: morphological criteria (P < 0.01), Annexin V (control, 5.01 ± 0.61; ETBP, 10.60 ± 1.98%, P = 0.0005), and TUNEL (control, 12.5 ± 2.6; ETBP, 20.56 ± 3.06%, P = 0.001; ETB, 30.69 ± 5.05%, P = 0.001). Increased expression of apoptosis was accompanied by increased expression of Fas (control, 20.15 ± 5.28; ETBP, 43.51 ± 5.6%, P = 0.03; ETB, 47.16 ± 5.54%, P = 0.01), Fas ligand (control, 5.64 ± 2.38; ETBP, 11.66 ± 3.65%, P = 0.04; ETB, 16.39 ± 5.05%, P = 0.02) and p53 products (control, 9.22 ± 3.44; ETBP, 22.82 ± 5.72%, P = 0.01; ETB, 24.60 ± 5.20%, P = 0.01). Treatment of ETB-leukocyte cultures with anti-human Fas-L exhibited 2.2-fold lower apoptosis expression. Treatment with E64 significantly abrogated the apoptotic effect of ETB. There was no increment on leukocyte oxidative metabolism. Mononuclear leukocytes also showed elevated levels of proliferation when treated with different concentrations (from 50 to 6.2 μg/mL) of streptococcal proteins (Stimulation index ranging: ETBP, 5.6 ± 1.9 to 6.4 ± 1.9; ETB, 9.9 ± 2.8 to 13.9 ± 3.8). Conclusions. These results delineate an additional pathway for the pathogenesis of APSGN related to the role of cationic streptococcal ETB or ETBP on the induction of apoptosis and proliferation during the course of the disease.

AB - Background. Previous reports have shown the presence of erythrogenic toxin type B (ETB), apoptosis, proliferation, and leukocyte infiltration in biopsies from patients with acute poststreptococcal glomerulonephritis (APSGN). Methods. Attempting to correlate the apoptotic and proliferative events with the interaction of ETB or its precursor (ETBP) with leukocytes, mononuclear leukocytes from 12 healthy subjects were cultured with ETB or ETBP to analyze the levels of apoptosis, proliferation, expression of modulatory apoptosis gene products, and oxidative metabolism. After four days of incubation, cells were assessed for apoptosis by morphological criteria, annexin V assay, and terminal deoxy transferase uridine triphosphate nick end-labeling (TUNEL) assay. The expression of regulatory apoptosis genes was assessed by relevant monoclonal antibodies; proliferation was by incorporation of radioactive thymidine; and oxidative metabolism was by oxidation of 2′,7′-dichlorofuorescein diacetate to 2′,7′-dichlorofuorescein. Neutralization of Fas-L and cysteine protease activity of ETB were performed by incubation of ETB-treated leukocyte cultures with anti-human Fas-L mAb or with E64, respectively. Results. Elevated levels of apoptosis in ETBP/ETB-treated leukocytes were found when compared with controls: morphological criteria (P < 0.01), Annexin V (control, 5.01 ± 0.61; ETBP, 10.60 ± 1.98%, P = 0.0005), and TUNEL (control, 12.5 ± 2.6; ETBP, 20.56 ± 3.06%, P = 0.001; ETB, 30.69 ± 5.05%, P = 0.001). Increased expression of apoptosis was accompanied by increased expression of Fas (control, 20.15 ± 5.28; ETBP, 43.51 ± 5.6%, P = 0.03; ETB, 47.16 ± 5.54%, P = 0.01), Fas ligand (control, 5.64 ± 2.38; ETBP, 11.66 ± 3.65%, P = 0.04; ETB, 16.39 ± 5.05%, P = 0.02) and p53 products (control, 9.22 ± 3.44; ETBP, 22.82 ± 5.72%, P = 0.01; ETB, 24.60 ± 5.20%, P = 0.01). Treatment of ETB-leukocyte cultures with anti-human Fas-L exhibited 2.2-fold lower apoptosis expression. Treatment with E64 significantly abrogated the apoptotic effect of ETB. There was no increment on leukocyte oxidative metabolism. Mononuclear leukocytes also showed elevated levels of proliferation when treated with different concentrations (from 50 to 6.2 μg/mL) of streptococcal proteins (Stimulation index ranging: ETBP, 5.6 ± 1.9 to 6.4 ± 1.9; ETB, 9.9 ± 2.8 to 13.9 ± 3.8). Conclusions. These results delineate an additional pathway for the pathogenesis of APSGN related to the role of cationic streptococcal ETB or ETBP on the induction of apoptosis and proliferation during the course of the disease.

KW - Cell death

KW - DNA fragmentation

KW - Glomerulonephritis

KW - Phosphatidylserine

KW - Streptococcal proteinase

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