Diversity of antibodies to cross‐reacting nitrophenyl haptens in inbred mice

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Abstract
The use of cross‐reacting antigens to stimulate antibody responses of limited heterogeneity is described. The cross‐reacting haptens 2,4dinitrophenyl (DNP) and 2,4,6trinitrophenyl (TNP), coupled to ovalbumin or bovine IgG, were employed as immunogens in inbred CBA/J and C3H/He mice. Sera from mice primed with DNP‐protein and subsequently boosted with TNP coupled to the same carrier were examined by isoelectric focusing techniques. The anti‐DNP antibodies in these sera were shown to consist of the products of a small number (usually less than 4) of clones.The frequency of occurrence of anti‐DNP antibodies that were indistinguishable by isoelectric focusing techniques was estimated. In 2400 comparisons between antibodies in sera from 140 individual mice, a small number (5 to 7) of indistinguishable antibody pairs was found. It was estimated from these results that not less than about 500 different sorts of antibodies to DNP, strongly cross‐reacting with TNP, can be made by inbred mice immunized with these cross‐reacting antigens. The problem of natural selection for the ability to produce such a large variety of antibodies is discussed.