Four groups of hysterectomy-derived and colostrum-deprived pigs were intranasally inoculated with an Actinobacillus pleuropneumoniae biotype 1-serotype 2 strain (producing RTX toxins ApxII and ApxIII. 6 pigs), an A. pleuropneumoniae biotype 1-serotype 10 strain (producing ApxI. 5 pigs), an A. pleuropneumoniae biotype 2-serotype 2 strain (producing ApxII, 5 pigs) or saline (controls, 7 pigs). All pigs were exposed to A. pleuropneumoniae biotype 1-serotype 2 endobronchial challenge. After challenge, severe clinical signs were observed in all control pigs, one pig immunized with the A. pleuropneumoniae biotype 1-serotype 10 strain and two pigs immunized with the A. pleuropneumoniae biotype 2-serotype 2 strain. These pigs died within 36 h after challenge and 20 to 50% of the lungs were macroscopically affected. In the other pigs, clinical signs were mild or absent and no or only small, focal lung lesions were observed when euthanized at 48 h after challenge. At the time challenge neutralizing antibodies against ApxI only. ApxII only and both ApxII and III were present in sera of pigs immunized with the A. pleuropneumoniae biotype 1-serotype 10 strain, the A. pleuropneumoniae biotype 2-serotype 2 strain and the A. pleuropneumoniae biotype 1-serotype 2 strain, respectively. These results indicate that immune mechanisms other than Apx neutralizing antibodies were involved in partial cross-protection of pigs immunized against A. pleuropneumoniae biotype 1-serotype 10 and challenged with the A. pleuropneumoniae biotype 1-serotype 2.