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. 1997 Nov;56(11):671–676. doi: 10.1136/ard.56.11.671

Invasion and persistence of Salmonella in human fibroblasts positive or negative for endogenous HLA B27

H Huppertz 1, J Heesemann 1
PMCID: PMC1752276  PMID: 9462170

Abstract

OBJECTIVE—Analysis of the interaction of enteropathogenic bacteria with HLA B27 transfected murine fibroblasts showed a specific influence of HLA B27 on microbial invasiveness. This possible novel mechanism for the action of HLA B27 should be verified by using endogenous HLA B27 positive and negative human fibroblasts as a model for the direct interaction of arthritogenic bacteria and host cells.
METHODS—Fibroblasts were obtained from healthy donors positive or negative for HLA B27; cultivated as monolayers; and infected with Salmonella enterica serovar enteritidis.
RESULTS—Invasion and survival of bacteria in human cells was not influenced by the presence of HLA B27. Enhancement of HLA class I molecule expression by treatment of cultures with interferon gamma decreased invasion and survival of bacteria in both HLA B27 positive and negative cells. After disappearance of live bacteria lipopolysaccharide antigens persisted within cells.
CONCLUSION—Endogenous HLA B27 does not modulate the direct interaction of Salmonella with human cells. Non-professional phagocytes are able to limit bacterial survival in cells, and interferon gamma accelerates killing of bacteria, but arthritogenic antigens persist after disappearance of live bacteria.



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Figure 1  .

Figure 1  

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Influence of HLA B27 on invasion and survival of Salmonella enterica serotype enteritidis in 26 primary human fibroblast lines, six of which were HLA B27 positive and 20 HLA B27 negative. (A) Number of salmonellas per 100 cells after the infection (invasion) and one and seven days later (survival). Median (range) in HLA B27 positive/ negative cells were: day 0: 210 (30-268) / 251 (68-348); day 1: 505 (282-1280) / 794 (282-1910); day 7: 1 (1-6) / 2 (1-4). (B) Colony forming units per monolayer (CFU/ml) after the infection and one and seven days later. Median (range) in HLA B27 positive / negative cells were: day 0: 4.6 × 104 (2.6-11 × 104) / 7.4 × 104 (1.7-17 × 104); day 1: 9.5 × 105 (2.7-23 × 105) / 12.0 × 105 (2.5-27 × 105); day 7: 30 × 102 (3.1-160 × 102) / 22 × 102 (6.0 -150 × 102). Comparing the groups of HLA B27 positive and negative cell lines for the number of salmonellas per 100 cells and the CFU/ml at the three time points by U test did not yield any significant differences.

Figure 2  .

Figure 2  

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Persistent infection of Salmonella enterica serotype enteritidis in primary human fibroblasts. Three cell lines are HLA B27 positive, two lines HLA B27 negative.

Figure 3  .

Figure 3  

Figure 3  

Figure 3  

Figure 3  

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Salmonella antigen in primary human fibroblasts by indirect immunofluorescence staining for Salmonella lipopolysaccharide antigens directly after infection and two hours of antibiotic treatment (A) and seven (B) and 21 days later (C). Initially intact bacterial rods prevail. After seven days aggregates of cytoplasmic antigen deposits can be seen (arrowhead). After 21 days, at a time when the culture had become sterile, bacterial rods can no longer be identified, but finely dispersed antigen deposit can be seen in the cells. For control a parallel culture was stained with normal rabbit serum after 21 days of infection (D); to show cell morphology exposure time was doubled in comparison with fig 3C. Staining with the normal rabbit serum shows no green colour, but only faint yellow non-specific background staining.

Figure 4  .

Figure 4  

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Effect of treatment with interferon gamma on the survival of Salmonella in primary human fibroblasts. Seven cell lines were infected with Salmonella enterica and then treated with 1000 IU/ml of interferon gamma or left untreated. The three HLA B27 positive cell lines are displayed with square symbols, the four HLA B27 negative cell lines with round symbols. Using ANOVA to compare treated and untreated cells treatment effect (p<0.001) and interaction (p<0.0001) were significantly different.

Figure 5  .

Figure 5  

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Effect of pre-treatment with interferon gamma on the invasion of Salmonella in primary human fibroblasts. Seven cell lines were treated with 1000 IU/ml of interferon gamma for 48 hours or left untreated and then infected with Salmonella enterica. The three HLA B27 positive cell lines are displayed with square symbols, the four HLA B27 negative cell lines with round symbols. Using ANOVA to compare treated and untreated cells treatment effect was highly significant (p<0.000005), but interaction was not, confirming the significant influence of interferon gamma on invasion.

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