Update: Is It the Blood of Louis XVI?

Statue of King Henry IV at Pont Neuf in Paris

Statue of King Henry IV at Pont Neuf in Paris

In an earlier blog entry, I wrote about the ill-fated Louis XVI, the French king who was famously beheaded along with his wife, Marie-Antoinette, during the French Revolution in 1793. Witnesses to the execution dipped handkerchiefs in the king’s blood and kept them as souvenirs of the common people’s rebellion. In 2010, scientists published the presumptive DNA profile of the king, obtained from one of these bloody handkerchiefs (1). Shortly after this profile was published, doubters surfaced, arguing that scientists could not say with certainty that the blood was that of Louis XVI. Clearly, more work was needed to identify the source of the blood. Recently, additional work was published (2,3). The most recent data (3) were presented at the International Symposium on Human Identification; these newest data cast doubt on the identification of the remains of not one king, but two.

Those remains have been attributed to King Henry IV and his great-great-great-great-great grandson Louis XVI, who both ruled over France for many years (1589–1610 and 1774–1791, respectively). As I mentioned previously, Louis XVI was not a popular king, and his mismanagement of French finances helped spawn the French Revolution and led to his swift but gruesome death at the guillotine. Initially, Henry IV also was not popular with many of his subjects. In the late 16th century, then-King Henry III’s reluctant acknowledgement of Henry IV as the legitimate successor to the French throne kicked off a civil war known as the Wars of Religion or the War of Three Henrys. Henry of Navarre, as he was known prior to his ascension to the French throne as Henry IV, was baptized as a Catholic but raised as a Protestant, and his claim to the throne was strongly opposed by the Catholic League, who funded and waged many battles against his forces. However, after many years of war and Henry III’s death, Henry IV was crowned king, renounced his Protestantism and issued the Edict of Nantes, which ushered in a period of religious tolerance. Henry IV’s reign was also a time of great prosperity, in part due to his emphasis on improving education and his efforts to promote trade and commerce. He became one of France’s best loved rulers and was the founding monarch of the House of Bourbon, which included all subsequent French kings until the monarchy was abolished in 1792. Despite his reputation as “Good King Henry”, his grave at Basilica of St Denis, along with many other royal graves, was desecrated during the French Revolution, and his mummified head was stolen. The presumptive head resurfaced in the 1900s at an auction in Paris, where it was bought by a private collector. It was these remains, the presumptive head of Henry IV and the blood on the handkerchief, that were used to perform additional DNA testing to try to provide more definitive identifications.

The original study of the bloody handkerchief (1) examined DNA from the Y chromosome, which is inherited through the paternal lineage, and mitochondrial DNA (mtDNA), which is inherited through the maternal lineage, to generate the presumptive DNA profile of Louis XVI, which included hypervariable regions 1 and 2 sequences, a limited Y-STR profile and some autosomal STR markers. Unfortunately, no familial reference samples were analyzed at the time to try to confirm the provenance of the blood. To generate more complete profiles and strengthen the argument that the blood belonged to Louis XVI, members of the same laboratory performed additional DNA analyses and compared the results to a DNA profile from a mummified head attributed to Henry IV through a combination of anthropological, pathological, radiological, historical and genetic data (4). Henry IV and Louis XVI should have identical mtDNA sequences and Y-STR profiles (barring any DNA mutations) because they were both descended from Anna of Habsburg through the maternal lineage and Louis XVI was descended from Henry IV through a direct paternal lineage. This new analysis provided partial Y-STR profiles and HV1 sequences that were consistent with related males separated by seven generations (2).

End of story, right? Wrong. Skeptics argued that the “DNA match” between the blood and head could have been by chance because the DNA profile from the head was very limited (5). These scientists performed their own DNA testing, and those results were just published October 9 in the European Journal of Human Genetics (3).

In this most recent study, scientists re-analyzed DNA from the remains plus three living descendants of Henry IV. They examined the Y chromosome and were able to generate the most complete DNA profiles to date, although the head again yielded very limited genetic data due to its age. As direct descendants through the paternal lineage, the three members of the House of Bourbon should share the same (or very similar) Y-STR profile with both Henry IV and Louis XVI. However, that did not seem to be the case: The Y-STR analyses revealed numerous differences between the true Y-STR haplotype of the House of Bourbon and the Y-STR haplotypes generated from the royal remains, supporting their hypothesis that the blood and mummified head are not those of Louis XVI and Henry IV, respectively.

The controversy doesn’t end there though. One of the researchers who helped identify the mummified head points out possible infidelity in the French monarchy. A nonpaternity event at keys points in the royal lineage would explain why there was no Y-STR “match” between living members of the House of Bourbon and the head and blood samples. This researcher and his colleagues plan to publish additional evidence that the head was properly identified: a three-dimensional analysis of the head and Henry IV’s death mask, which shows an exact match.

Will that be the definitive answer to the source of the blood and mummified head?

For some reason, I doubt it.

References

  1. Lalueza-Fox, C. et al. (2010) Genetic analysis of the presumptive blood from Louis XVI, king of France. Forensic Sci. Int. Genet. 5, 459–63. PMID 20940110.
  2. Charlier, P. et al. (2013) Genetic comparison of the head of Henri IV and the presumptive blood from Louis XVI (both Kings of France). Forensic Sci. Int. 226, 38–40. PMID 23283403
  3. Larmuseau, M.H. et al. (2013). Genetic genealogy reveals true Y haplogroup of House of Bourbon contradicting recent identification of the presumed remains of two French Kings. Eur. J. Hum. Genet. DOI: 10.1038/ejhg.2013.211
  4. Charlier, P. et al. (2010) Multidisciplinary medical identification of a French king’s head (Henri IV). Br. Med. J. 341, c6805. PMID 21156748.
  5. Fornaciari G. (2011) A French king’s head. Was it Henri IV’s head? Br. Med. J. 342, d293.
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Terri Sundquist

Terri has worked as a Scientific Communications Specialist at Promega Corporation for more than 13 years, and prior to that, spent more than 5 years solving problems and answering questions as a Promega Technical Services Scientist. She graduated with B.S. degrees in Chemistry and Biology at the University of Wisconsin—River Falls, then earned her M.S. in Molecular Biology from the Mayo Graduate School in Rochester Minnesota.

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