In the five-year study, California, Connecticut, Massachusetts, New York and Pennsylvania showed the most negative tweets of any states in the US.
Regions with high affluence or a large number of new moms were most likely to be hotbeds of anti-vaccine Twitter users, the study found.
"The debate online is far from over. There is still a very vocal group of people out there who are opposed to vaccines," said Chris Vargo, assistant professor at University of Colorado Boulder in the US.
For the study, published in the journal Social Science and Medicine, researchers created a machine-learning algorithm to examine more than a half-million tweets from around the country between 2009 and 2015.
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To make the sample a manageable size, they looked only at tweets that referred to both autism spectrum disorder and vaccines.
For two decades anti-vaccine activists have suggested that certain vaccines can lead to autism, often referring to a 1998 study of 12 children, published in the Lancet, which suggested that the measles, mumps, and rubella (MMR) vaccine predisposed youth to developmental disorders.
"Time and time again researchers have tried to substantiate this idea that there is a link between autism and vaccines but they have not been able to," said Theodore Tomeny, autism researcher with University of Alabama in the US.
"Unfortunately the idea is still very much out there, being promoted by a vocal minority online. That's problematic because often only one side of the story is being told," Tomeny said.
The researchers note that recent outbreaks of previously eradicated, vaccine-preventable diseases like measles and pertussis have been linked to refusal to vaccinate and anti- immunization-related beliefs.
Between 2010 and 2015, anti-vaccine tweets became, overall, more common nationwide. As the number of households that made over USD 200,000 annually increased or the number of women who had delivered a baby in the past 12 months increased, so did the amount of anti-vaccine tweets in a particular region.
Vargo stressed that he does not see Twitter posts as a representative sample of overall public opinion, but rather a pulse of the level of anti-vaccine activism in an area.
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