CD4(+) T cells specific to a glomerular basement membrane antigen mediate glomerulonephritis - PubMed (original) (raw)

CD4(+) T cells specific to a glomerular basement membrane antigen mediate glomerulonephritis

Jean Wu et al. J Clin Invest. 2002 Feb.

Abstract

Ab-mediated mechanisms have been considered the major causes of glomerulonephritis (GN). However, recent studies suggest that T cells may be more important in mediating GN. To investigate the effects of antigen-specific CD4(+) T cells, we generated Th1 cell lines specific for this antigen from rats that had been immunized with a recombinant form of the glomerular basement membrane (GBM) antigen, Col4alpha3NC1. Upon the transfer of in vitro-activated T cell lines to pertussis toxin-primed, naive syngeneic rats, the recipients developed severe proteinuria/albuminuria, which plateaued after approximately 35 days. Although no IgG binding to GBM or C3 deposition could be detected by immunofluorescence, five out of eleven rats exhibited severe GN, as judged by the formation of characteristic crescent-shaped lesions in the glomeruli, whereas the others exhibited modest GN. Thus Col4alpha3NC1-specific T cells directly initiated glomerular injury in the recipients. One notable difference from GN induced by active immunization was a T cell infiltration in the renal interstitium, which affected some tubules. We therefore injected fluorescence-labeled Col4alpha3NC1-specific into naive rats, and we found that they were enriched 4.5-fold in the kidney cortex relative to nonspecific control T cells 24 hours later. Many of the T cells were located in the Bowman's space and had a flattened shape, suggesting that the primary target for the T cells was in or adjacent to the Bowman's capsule.

PubMed Disclaimer

Figures

Figure 1

Figure 1

Antigen specificities of a T cell line generated from rCol4α3NC1-immunized rat. (a) An rCol4α3NC1 T cell line (filled circles) or T cells from CFA-immunized rat (open circles) were incubated with different concentrations of rCol4α3NC1 (x axis) in the presence of irradiated syngeneic thymocytes. The proliferation of T cells was measured as uptake of 3H (Δcpm, y axis). (b) T cells’ responses to digested rat GBM.

Figure 2

Figure 2

Cellular composition of rCol4α3NC1-specific T cell lines. (a) Flow cytometry analysis of whole lymphocytes from rCol4α3NC1-immunized rats (top panel) and a T cell line (bottom panel). (b) Rat IgG concentration in the supernatants of two Col4α3NC1 T cell lines (open circles and squares) and one control CFA T cell line (filled circles) collected after each stimulation (x axis). Ab activities to rCol4α3NC1 in the supernatants from a Col4α3NC1 T cell line, determined by Western blot analysis, are shown (inset).

Figure 3

Figure 3

Cytokine profiles in an rCol4α3NC1-specific T cell line. (a) RT-PCR detection of rat INF-γ/IL-4 expressions at rest (R) or after rCol4α3NC1 stimulation (S). DNA size markers are shown at the left. (b) Flow cytometry analyses on cells from a Col4α3NC1-specific T cell line at rest (left panel) or stimulated by mitogens (right panel) after intracellular stain with anti–rat IFN-γ and IL-4 Ab’s.

Figure 4

Figure 4

Proteinuria and albuminuria development in the rCol4α3NC1-specific T cell recipients. (a) Urine samples were collected as indicated (x axis) in 11 T cell recipients, and proteinuria was measured (y axis). Filled symbols, rats with severe GN; open symbols, modest GN; dashed lines, normal kidney. (b) Urine samples from a representative T cell recipient (T cell) with severe GN and a rat immunized with rCol4α3NC1 (rCol) were analyzed by SDS-PAGE. Arrows at right indicate serum albumin.

Figure 5

Figure 5

Development of GN in the rCol4α3NC1-specific T cell recipients. (a) Histology shows a glomerulus with fibrin deposition and crescent formation. Adjacent to the glomerulus is a mononuclear infiltrate. Focal tubules are dilated and contain intensely eosinophilic material consistent with protein. (b) Immunoperoxidase staining shows CD3+ T cells (brown in color) in mononuclear cell infiltrate surrounding a glomerulus with crescentic lesion. (c) Immunofluorescence stain show CD11b/c+ monocytes/macrophages (red) clustered in the Bowman’s space of an affected glomerulus. GBM and TBM are counterstained green by FITC-labeled SR-6. Arrowheads outline the basement membrane of the Bowman’s capsule.

Figure 6

Figure 6

EM micrograph shows segmental effacement of visceral epithelial cell (podocytes) foot processes (arrows) in an rCol4α3NC1-specific T cell recipient.

Figure 7

Figure 7

Direct immunofluorescence of Ab on serial frozen sections of kidney from an rCol4α3NC1 T cell recipient. (a) Section stained with FITC-labeled anti-rat IgM Ab. Crescent-shaped fluorescence staining is visible. (b) Section stained with Ab to rat IgG (negative). (c) Counterstaining by methylene blue illustrates fibrin in a crescentic lesion in glomerulus.

Figure 8

Figure 8

Tracing CFDA-SE–labeled T cells in kidney cortex by fluorescent microscopy. (a and b) rCol4α3NC1-specific T cells or CFA T cells were transferred into naive recipients. The labeled cells were counted in serial frozen sections of the kidney cortex 24 hours later, and labeled cell density was calculated (a). Elongated cells are expressed as a percentage of all labeled cells (b). (c) Fluorescent microscopy illustrates an elongated rCol4α3NC1-specific cell (green) in the Bowman’s capsule. GBM is stained red by PE-labeled SR-13. The tubules (T) with background are visible. Inset: high power of two elongated, labeled T cells.

Comment in

Similar articles

Cited by

References

    1. Wilson, C.B. 1996. Renal response to immunologic glomerular injury. In The kidney. Volume 2. B.M. Brennen, editor. W.B. Saunders Co. Philadelphia, Pennsylvania, USA. 1263–1349.
    1. Unaue ER, Dixon FJ. Experimental allergic glomerulonephritis induced in the rabbit with heterologous renal antigens. J Exp Med. 1967;125:149–163. - PMC - PubMed
    1. Kalluri R, Gattone VH, Jr, Noelken ME, Hudson BG. The α3(IV) chain of type IV collagen induces autoimmune Goodpasture syndrome. Proc Natl Acad Sci USA. 1994;91:6201–6205. - PMC - PubMed
    1. Theofilopoulos AN, Dixon FJ. Immune complexes in human diseases: a review. Am J Pathol. 1980;100:529–594. - PMC - PubMed
    1. Wang Y, et al. Amelioration of lupus-like autoimmune disease in NZB/W F1 mice after treatment with a blocking monoclonal antibody specific for complement component C5. Proc Natl Acad Sci USA. 1966;93:8563–8568. - PMC - PubMed

Publication types

MeSH terms

Substances

LinkOut - more resources